CN105025905A - Methods for use of sex sorted semen to improve genetic management in swine - Google Patents
Methods for use of sex sorted semen to improve genetic management in swine Download PDFInfo
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- CN105025905A CN105025905A CN201380074208.1A CN201380074208A CN105025905A CN 105025905 A CN105025905 A CN 105025905A CN 201380074208 A CN201380074208 A CN 201380074208A CN 105025905 A CN105025905 A CN 105025905A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0278—Physical preservation processes
- A01N1/0284—Temperature processes, i.e. using a designated change in temperature over time
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D19/00—Instruments or methods for reproduction or fertilisation
- A61D19/02—Instruments or methods for reproduction or fertilisation for artificial insemination
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/52—Sperm; Prostate; Seminal fluid; Leydig cells of testes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
Abstract
The invention relates to methods of increasing genetic merit of swine using sex sorted semen by establishing a plurality of mating subtypes for a line of swine, and determining a percentage of progeny that are male for each of the mating subtypes, or a percentage of progeny that are female for each of the mating subtypes, that would result, relative to a control, in an increase in genetic merit in the line.
Description
This application claims rights and interests and the priority of the U. S. application numbers 13/840,598 submitted on March 15th, 2013, the priority of its U. S. application requiring on June 6th, 2012 to submit to numbers 61/656,446.
Invention field
The present invention relates to the method using the sperm of the sex sorting of the pure lines boar from the pig system be in copulation to increase the genetic worth of pig in the market pig level of production.The improvement of the genetic management of this strain is the result of following item: the increase of the genetic progress 1) in this strain and/or 2) hereditary improvement (that is, the genetic worth of commercial product is closer to the genetic worth at hereditary core place) of transmitting.
Background
Current pig produces can by multistage pyramid representation, a certain offspring being wherein in each grade in next even lower level not for breeding.Pyramidal top be hereditary core (GN).Immediately rank is from top to bottom daughter nucleus (DN) substantially, multiplier or propagation unit, finally for business level, usually comprise the commercial farm of wherein production slaughter pig respectively.
Typically, the genetic progress of a strain occurs in and is in the pure lines colony of hereditary core.GN animal will have and will be in pyramidal more low-level relatives, and purebred also have cenospecies.The trait data collected from these relatives contributes to the genetic worth estimating GN animal.Within the strain being in GN, once selected, usually make the follow-on parent of generation random mating each other, avoid the copulation between closely related individual simultaneously, target is to increase follow-on genetic worth.Increase on follow-on genetic worth contributes to genetic progress.Under this background, the increase on genetic worth means for given character or a set of character, and the individuality in the successive generation will show desired character or a set of character stronger than its parent.About bad character, the increase on genetic worth means that the individuality in the successive generation will show the character strong not as its parent or a set of character.
Hereditary change, comprises desired hereditary change (that is, annual genetic progress), the difference between all filial generations that (" dG ") can be measured as in a year birth and the average genetic level of all filial generations of being born at next year.This difference is the result than the average genetic worth of all selection candidates (can be used for the animal selected) with the selected parent of higher genetic worth.Under ideal conditions, this depends on the heritability (h of character
2) and selected parent and select candidate mean apparent between difference.Heritability (the h of character
2) be relative to observable difference (phenotypic variance, the σ in the character between intragroup individuality caused by additive inheritance (A) of environment (E) difference
2 p) ratio (h
2=σ
2 a/ σ
2 p=σ
2 a/ (σ
2 a+ σ
2 e)).Difference between selected parent and the mean apparent of all selection candidates (its selected parent is a subset) be also called as select poor.
Annual genetic progress is selected male and selected female prepotent result.This is expressed as following equation:
DG={ (R
iH* i)
male+ (R
iH* i)
female* σ
h/ (L
male+ L
female),
Wherein, for male or female parent, the accuracy that R=selects, i=selection intensity, σ
h=hereditary variation, and L=generation inteval.
These character (1 breeding objective (H=v merged to genetic worth (g) m) that the needs of economic worth (v) weighting by character produce by H=
1g
1+ v
2g
2+ ... + v
mg
m).If select be for larger phenotypic number, then this economic worth is positive, and if select be for less phenotypic number, this this economic worth is negative.
The index that all trait information that are individual and relatives merge by I=, and be the best estimate of the H-number to this individuality.
When remove non-hereditary effect (such as, by by each track record and the same period group meansigma methods compare) and when using the information from relatives except this animal self, selection is more effective.This realizes by using such as Multi-trait BLUP method to calculate estimated breeding value (EBV).Envirment factor such as HYS (field-year-Ji) is used for correcting environmental effect in a model, and simultaneously by using relational matrix the information from relatives to be included.More multiple characters information from more relatives causes the accuracy (R of higher EBV
iH).
In large group, selection intensity depends on and tests how many animals and have selected how many animals-when other conditions are identical, and the ratio of selection is lower, then selection intensity is higher and genetic progress is larger.Therefore, in order to make genetic progress maximize, people should based on EBV by all test animal classifications, and then select to maintain this strain, breeding and/or drove size and avoid the minimal amount of top boar required for inbreeding problem and sow.This guarantees that the average EBV of selected animal is in fact higher than the average EBV of all test animals.Especially by utilizing artificial insemination to test (AI), the boar that people need Selection radio gilt less, and for male selection intensity higher than female.
Generation inteval for male (or female) is mean age of the male parent (or female parent) when offspring born.Generally speaking, sow produces the little son more than a tire at GN place, and female L tends to larger than male L.
The annual rate of genetic progress depends on the generation inteval and depends on that parent selects the advantage of the EBV of candidate compared to these.Generally speaking, male more for the contribution of annual genetic progress than female.
The example of the important character in pig industry is feed efficiency, that is, feeding quality is being converted into the measuring of efficiency (also referred to as food conversion or feed-weight ratio) of body quality of increase and the average daily gain of animal by animal, that is, average weight increase every day.With the character of different Unit-Measure (such as, the number of pig, the pound number of every day, inch, etc.) on all world markets, not there is equal Economic Importance, and be not affected (that is, different coefficient of heredity) with identical degree in heredity.In general, the production traits of such as feed efficiency and average daily gain and so on has high heritability.By contrast, such as the reproduction trait of fertility and tire litter size and so on has low heritability usually.
Operation cost for the genetic progress rate increased in pig industry in strain and reduction breeding and business pig farm also exists needs.
Summary of the invention
Some embodiment of the present invention comprises a kind of method increasing the genetic worth of pig, and the method comprises the following steps: set up the multiple copulation hypotypes for a strain; The percentage ratio determining in these copulation hypotypes each to be each in the percentage ratio of male filial generation or these copulation hypotypes be female filial generation, these filial generations will cause the increase on genetic worth relative to contrast; Divided by spermatid sample from the male hog be in one of these copulation hypotypes and elect one or more spermatid subgroup as, the most of spermatids in one of them spermatid subgroup are with X chromosome or Y chromosome; The one or more female pigs inseminations be in one of these copulation hypotypes are made, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast with the spermatid of this subgroup; And from this one or more female pigs production filial generation.In an other embodiment, being the percentage ratio of male filial generation for each in these copulation hypotypes or being the percentage ratio of female filial generation for each in these copulation hypotypes, causing relative to impinging upon the increase on genetic worth, the inbreeding in this strain not increasing.In an other embodiment, the step of this insemination can by make by this spermatid subgroup from the one or more female pigs be in one of these copulation hypotypes one or more ovums in vivo or the step of in vitro fertilization replace, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast.In certain embodiments of the present invention, this strain, this male, this one or more female and/or filial generation, can belong to or be the member of hereditary core, daughter nucleus or multiplier.In other embodiments of the invention, any or all of above-mentioned steps can perform as a part for a procedure of breeding.Should be appreciated that some embodiment of the present invention comprises the one or more of above-mentioned steps.
Be to be understood that, in certain embodiments of the present invention, in this spermatid subgroup at least about spermatid of 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% with X chromosome or Y chromosome.
In certain embodiments of the present invention, the genetic worth of pig or strain can be quantitative or the function of genome EBV, based on quantitative or genome EBV or determined by quantitative or genome EBV.In other embodiments of the invention, genetic worth can be one or more character function, determine based on one or more character or by one or more character, described character includes but not limited to, fertility, tire litter size, milk yield, feed efficiency, average daily gain and carcass lean meat percentage, and for the genetic marker of this type of character.In an other embodiment, genetic worth can be need sex sorting and/or freezing spermatid ability function, based on need sex sorting and/or freezing spermatid ability or determined by the ability of sex sorting and/or freezing spermatid of needing, its based on sorting and/or freezing after the vigor of spermatid, fertilizability and/or energy and the genetic marker for this character.
In certain embodiments of the present invention, strain can comprise a paternal line or one maternal.Of the present invention in some, a strain can comprise a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35% or 15% to 30%.In other respects, strain can comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30% or 10% to 25%.In other again, be the percentage ratio of male filial generation for each in these copulation hypotypes or be female filial generation for each in these copulation hypotypes percentage ratio can between about 0 to 10% or between about 90% to 100%, and the filial generation produced can be the member of daughter nucleus or multiplier.
In certain embodiments of the present invention, the filial generation of about 50% is male in contrast.In other embodiments, the female pigs insemination of copulation is made all to need with the non-sorting sperms sample in contrast.
More of the present invention in, the classification of the male hog in one or more copulation hypotype or grade can define by one or more feature (comprising genetic worth or age).In other respects, the classification of the female pigs in one or more copulation hypotype or grade can define by one or more feature (comprising genetic worth or parity).In certain embodiments, each of these copulation hypotypes in multiple copulation hypotype can comprise an only male hog from this strain and/or an only female pigs.In some embodiments of the invention, a copulation hypotype can be made up of one or more subgroup and/or one or more female subgroup, one of them subgroup can be defined by one or more standard or based on one or more standard, described standard includes but not limited to, producing the function in pyramid, age, parity, genetic worth (such as, EBV) and genetic marker or sudden change.Some embodiments of the present invention comprise and will can be used for the male of copulation and/or female classification or be separated into the step of multiple subgroup, wherein these subgroups can be defined by one or more standard or based on one or more standard, described standard includes but not limited to, producing the function in pyramid, age, parity, genetic worth (such as, EBV) and genetic marker or sudden change.In an other embodiment, above-mentioned classification or separating step be carry out in a strain or as a part for a procedure of breeding or carry out for a step of the mating plan of a strain as foundation.In certain embodiments, male subgroup is defined by one or more standard or based on one or more standard, described standard includes but not limited to, is producing the function in pyramid, age, genetic worth (such as, EBV) and genetic marker or sudden change.In certain embodiments, female subgroup is defined by one or more standard or based on one or more standard, described standard includes but not limited to, is producing the function in pyramid, parity, age, genetic worth (such as, EBV) and genetic marker or sudden change.In some embodiments of the invention, one or more male subgroup can comprise one or more male.In other embodiments of the invention, one or more female subgroup can comprise one or more female.
Should be appreciated that in certain embodiments of the present invention, randomization method or Deterministic Methods or its combination can be used to determine the percentage ratio of the male and/or Female offspring increasing genetic worth.
In certain embodiments of the present invention, the genetic worth of pig or strain can be EBV function, determine based on EBV or by EBV.In other embodiments of the invention, genetic worth can be one or more character function, determine based on one or more character or by one or more character, described character includes but not limited to, fertility, tire litter size, milk yield, feed efficiency, average daily gain and carcass lean meat percentage, and for the genetic marker of this type of character.In an other embodiment, genetic worth can be need sex sorting and/or freezing spermatid ability function, based on need sex sorting and/or freezing spermatid ability or determined by the ability of sex sorting and/or freezing spermatid of needing, its based on sorting and/or freezing after the vigor of spermatid, fertilizability and/or energy and the genetic marker for this character.In certain embodiments of the present invention, by carrying out gene type to pig or embryo, the genetic worth of pig or strain can be evaluated.
In certain embodiments of the present invention, the seminal fluid of the sex sorting spermatid that can comprise sex sorting or the spermatid of sex sorting of once ejaculating and one or more other compositions.In other embodiments, the spermatid sample of sex sorting can comprise and wherein in this sample, causes its not fecundability spermatid sample by such as killing with the spermatid of X or Y.In other embodiments, the sex sorting method of spermatid sample or semen sample comprises wherein identified and be caused its not fecundability any method with the spermatid of X or Y in this sample.
Other aspects of the present invention are contained and use the spermatid of deep intrauterine catheter or the pin sex sorting inserted by the film of described female pigs to make to inseminate or breeding from the female pigs of described strain.Some in these embodiments contain and can be used for spermatid being placed to the known surgical in the reproductive tract of female pigs and No operation technology, comprise laparotomy (relating to the surgical operation of the large otch entering abdominal cavity through stomach wall).This embodiment is considered and is used 1x 10 altogether
9individual or less spermatid makes female pigs inseminate.
In other embodiments, deep intrauterine catheter can be adopted by spermatid sample administration in the distal portions (one or more cornua uteri or one or more uterotubal junction) of the reproductive tract of female pigs.In another aspect of the present invention, this deep intrauterine catheter is made up of an outer tube or sleeve pipe and an interior flexible probe.Of the present invention one other in, in this flexibility, probe comprises the wherein flexible innerduct that can pass of fluid or cell.In certain embodiments of the present invention, this outer tube and interior flexible probe can be made of plastics, and in other embodiments, they can be made up of the metal being configured to flexibility (as being in coil or spring configuration).In an other embodiment, this deep intrauterine catheter comprises the visual video camera in position or the endoscope that makes the distal portions of the deep intrauterine catheter in the reproductive tract of female pigs.In an alternative embodiment, radiography or fluoroscopy can be used to make this deep intrauterine catheter in the reproductive tract of female pigs visual.In another embodiment of the present invention, deep intrauterine catheter can be used embryo or conjugon to be inserted the distal portions (as from one or more cornua uteri or from one or more uterotubal junction) of the reproductive tract of female pigs or from it recall.
About with the insemination of deep intrauterine catheter, consider to use 1x 10 to female pigs
9individual spermatid or less dosage.Such spermatid can be the spermatid of sex sorting.In one embodiment of the invention, by spermatid (the such as 600x 10 of the sex sorting of a dosage
6individual, but can be more, if or be placed in optimum position at estrous Best Times and may be as few as 10x 10
6individual) one or two cornua uteri (such as, each angle 300x 10 of female pigs is administered to by deep intrauterine catheter
6individual spermatid) in.In other embodiments, dosage can at about 300x 10
6, about 150x 10
6, about 140x 10
6, about 100x 10
6, about 70x 10
6, about 50x 10
6, or about 5x 10
6any point change in the spermatid of individual sex sorting or less scope or within these scopes and can being applied in one or two cornua uteri of female pigs.
Above-mentioned dosage can be used with different volumes, include but not limited to, for every 150x 10
6the 5ml of individual spermatid, or in the volume of the cell of this identical number certain value in the scope of 5ml, 10ml, 15ml, 20ml, 25ml, 30ml or 100ml or between 5-10ml, 10-20ml, 20-30ml, 30-40ml, 40-50ml, 50-60ml, 60-70ml, 70-80ml, 80-90ml or 90-100ml.
Then the spermatid freezen protective of the sex sorting used in any embodiment of the present invention can be thawed, or alternately can utilize the spermatid of fresh (that is, not freezing) sex sorting.Above-mentioned dosage can also be administered in one or more uterotubal junctions of female pigs.
This embodiment of the present invention also contains that to use peritoneoscope the film by female pigs to be used the insertion of the pin of the spermatid sample of sex sorting visual.Can by be used for injecting spermatid sample pin and peritoneoscope and handle instrument (as pliers) by for laparoscopic surgery for typical minimal incision * is inserted in the abdominal part of female pigs.The present invention is also contained spermatid Sample Injection in the one or more positions within female reproductive tract.Be only citing, can this by spermatid Sample Injection in intrauterine one or more position of female pigs, comprise one or more cornua uteri, fallopian tube, ampulla, isthmus or uterotubal junction.In another embodiment of the present invention, embryo or zygote can be inserted in the reproductive tract of female pigs or from it via laparoscopy and recall.
About via laparoscopy insemination, consider to use 1x 10 to female pigs
9individual spermatid or less dosage.Such spermatid can be the spermatid of sex sorting.In one embodiment of the invention, can by laparoscopy by about 500x 10
6the dosage of the spermatid of individual or less sex sorting is expelled to (such as, 250x 10 in every oviductus lateralis in the one or both sides fallopian tube of female pigs
6individual spermatid); In other embodiments, can by about 10x 10
6, about 5x 10
6, about 3x 10
6, about 2.0x 10
6, about 1.2x 10
6, about 1x 10
6, or 0.6x 10
6within the scope of the spermatid of individual or less sex sorting, the dosage of (or any point within these scopes) is expelled in the one or both sides fallopian tube of female pigs.
In an other embodiment, by the spermatid injection of sex sorting in oviducal specific region, can include but not limited to, isthmus, ampulla and/or uterotubal junction.In certain embodiments, can by about 5x 10
6, about 2x 10
6, about 1x 10
6, about 600x 10
3, about 500x 10
3, about 300x 10
3, or about 150x 10
3in the scope of the spermatid of individual or less sex sorting, the dosage of (or any point within these scopes) is expelled in one or more regions of one-sided or bilateral salpingo.
In an other embodiment, with regard to regard to laparoscopy insemination, can the spermatid of different parts injection sex sorting in fallopian tube, be used in about 500x 10
3the dosage of the spermatid scope (or any point within this scope) of individual sex sorting is expelled in each ampulla, simultaneously by about 1x 10
6the spermatid injection of individual sex sorting is to each uterotubal junction; Or by about 1x 10
6the dosage of the spermatid of individual sex sorting is expelled in each ampulla, simultaneously by about 2x 10
6the spermatid injection of individual sex sorting is to each uterotubal junction; Or by about 5x 10
5the dosage of the spermatid of individual sex sorting is expelled in each ampulla, simultaneously by about 2x 10
6the spermatid injection of individual sex sorting is to each uterotubal junction; Or by about 5x 10
5the dosage of the spermatid of individual sex sorting is expelled in each ampulla, simultaneously by about 1x 10
6the spermatid injection of individual sex sorting is to each uterotubal junction.Above-mentioned dosage can be included in different volumes, such as, and every 1x 10
6(1,000,000) individual spermatid 100 μ l, or in the sperm of this identical number any volume in one of following volume or betwixt: 50 μ l, 100 μ l, 200 μ l, 300 μ l, 400 μ l or 500 μ l.
Another aspect of the present invention comprise embodiment that use discloses herein by use to female pigs one or more hormones or hormone analogs make to need the female pigs of inseminating estrus synchrony and/or induce its timing ovulation.In one embodiment, these one or more hormones or hormone analogs comprise PG600 and (comprise priatin [" PMSG "] and the human chorionic gonadotropin [" hCG "] of pregnant mare; Intervet (Intervet) company), OvuGel (via Intravaginal delivery system in the triptorelin acetate of controlled-release formulation; Gel medical science company (Gel MedSciences, Inc.)), horse chorionic-gonadotropin hormone (" eCG "), hCG, progestogen, altrenogest or RU-2267.
In an other embodiment of the present invention, one or more hormones described or hormone analogs are that the programmable device in the reproductive tract by being positioned over described female pigs is used.The programmable device considered herein can discharge one or more hormones described or hormone analogs in the mode of time controlled released, and breeder is except programming to the initial parameter for discharging one or more hormones described or hormone analogs, this device need not be monitored or any input is provided.In another embodiment of the present invention, by using the eCG of 1250 to 1500IU and used the hCG of 750IU after 72 to 80 hour, can estrus synchrony/timing ovulation in induction Female pig.In another embodiment, by using the PMSG of 400 to 2000IU and used 500 to the hCG of 1000IU after 72 to 83 hour, can rutting period in induction Female pig.
Other embodiments are further considered by checking that the follicle of described female pigs detects the ovulation in female pigs.In one particular embodiment of the present invention, the follicle of female pigs described in ultrasonic examination is used.In an other embodiment, 25-35 hour after hCG injection starts, by the ovary that existence every 3-5 hour check once described female pigs of Transrectal Ultrasound for preovulatory follicle.In an other embodiment of the present invention, within 2-3 hour after ultrasonic examination, select the female pigs demonstrating multiple preovulatory follicle for insemination.
Brief Description Of Drawings
Fig. 1 shows the production pyramid for the production of hybrid strain boar and parent gilt.
Fig. 2 shows and to cause the paternal and maternal increase of male filial generation on dG of complexity level increase in both and the reduction on percent of total when applying precise breeding.
Fig. 3 schematically illustrates the flow cytometer that spermatid sample can be used for divide and hank with one or more subgroups of X-or Y-chromosome.
Detailed description of the invention
These methods described herein by the genetic progress rate of an increase strain and/or to be reduced between GN and commodity production hereditary time apart from making propagation more cost-effective and/or have interests and add the genetic worth of pig in commercial level simultaneously.
" strain " refers to the pig with the common origin and similar recognition feature as used herein." pure lines " are equal to " strain " as used herein, and can use hereinafter, so that by purebred individuality or copulation and hybrid is individual or copulation distinguishes.
" procedure of breeding " comprises one or more strain exploitation program as used herein.
" market pig " refers to for its meat of commercial distribution by the pig butchered or the sow produced for the pig of its meat of commercial distribution.
" commercial farm " refers to the facility for holding market pig as used herein.
" multiplier " or " propagation unit " refers to male and one or more groups that are female pigs as used herein, comprise one or more strain, described product are a part for the propagation procedure of number for increasing the individuality with cumulative genetic worth, wherein for the individuality of pure lines or hybrid product is used as parent, for generations parent or the great grandparents of market pig.
" daughter nucleus " refers to the one or more groups for being sheerly the male of propagation and female pigs as used herein.
" hereditary core " refers to male and one or more groups that are female pigs as used herein, comprise one or more strain, described product are a part for the procedure of breeding of genetic worth for increasing this one or more strain, and can comprise or comprise the function of daughter nucleus.
As used herein " copulation hypotype " refer under list between the definition class of potential of copulation: 1) male definition classification, kind or type and available female; 2) available male and female definition classification, kind or type; Or 3) male definition classification, kind or type and female definition classification, kind or type.
" paternal line " refers to the strain of the production contributing to the parent boar used on commercial farm as used herein.
" maternal " refers to the strain of the production contributing to the parent gilt/sow used on commercial farm as used herein.
Produce in pyramid from hereditary core (" GN ") to each level of commodity production pig, for each copulation type, have selective male (boar and/or its seminal fluid) and female (gilt/sow and/or its ovum) for the production of filial generation.
At GN place, pure lines copulation occurs, creates male and female, and male and female being used for of the best being tested produces the next generation.Generally speaking (but such was the case with), GN male (or its seminal fluid) is only had for the heredity transmission to pyramidal even lower level other (daughter nucleus [" DN "] or multiplier [" M "]).
Usually, produce the parent for generations of slaughter pig at DN place, and produce the parent of slaughter pig in M level.In the example provided in FIG, for a kind of copulation type, a kind of sex in two kinds of sexes, is only had to need to produce in DN or M level.In business level other places, parent boar and parent gilt/sow produce slaughter pig.Producing the specific features separately depending on production system between pyramidal rank.In a closed drove pattern of trade, several rank can be found within a farm (structure).But with regard to copulation, usually still as shown in Figure 1ly process these copulation types.
With reference to figure 1, each letter-" A ", " B ", " C ", " D " and " E "-representative pure lines, wherein A and B is paternal, and C, D and E are maternal.In each square of Fig. 1, show copulation type or multiple copulation type (such as " A*A ", " D*E ", " C*DE " etc.) and desired piglets sex (" ♂ " and/or " ♀ ").The details (DN and M, wherein M generally includes parent boar M and parent gilt M) of propagation structure depend on breeding companies's (large or little), its consumer (large or little) and finished product (3,4 or 5-unit hybridize).Fig. 1 gives and produces a pyramidal example.The paternal line (such as, B) producing gilt and maternal (such as, the C) that produce boar can be seen.For each of these strains, superior test boar be moved to AI website and its seminal fluid for: 1) in the pure lines copulation at GN place, male and Female offspring and 2 desired by generation) pure lines copulation at DN/M place or hybrid copulation, the male or Female offspring desired by generation.
The increase of the genetic progress on GN is usually directed to the production of boar and gilt, test and selection.Under conventional cases, (that is, use unsorted seminal fluid), effectively have two kinds and select approach: produce paternal and maternal paternal line and produce paternal line and maternal maternal.Use the seminal fluid of sex sorting to set up use four kinds and select the chance of approach (namely, produce paternal paternal line [" SS "], produce maternal paternal line [" SD "], produce paternal maternal [" DS "] and produce maternal maternal [" DD "]), and in commercial level, increase genetic worth and genetic progress rate.The annual genetic progress rate be in a strain at GN selects accuracy, selection intensity, hereditary variation and the function of generation inteval:
dG＝{(R*i)
SS+(R*i)
SD+(R*i)
DS+(R*i)
DD}*σ
H/(L
SS+L
SD+L
DS+L
DD)，
Wherein,
R selects accuracy, wherein R substantially
sS=R
sD>=R
dS=R
dD
L is the generation inteval, wherein L
sSfor Damxung temper is for the mean age of connatae boar, L
sDfor the mean age when the connatae boar of Female offspring, L
dSfor Damxung temper is for the mean age of connatae sow, and L
dDfor the mean age when the connatae sow of Female offspring.
' i ' is for the selection intensity of each in four kinds of approach.
Approach is selected for available four kinds, people have an opportunity to select which concrete male parent or which concrete female parent within a colony to produce male offspring, and which concrete male parent or which concrete female parent produce female descendant within a colony.As described herein, the concept of " precise breeding " allows people accurately to find the male and female parent of the best for generation of particular sex, so that by using the copulation of the seminal fluid of sex sorting, test and select planning to increase genetic worth and genetic progress rate in commercial level.The use of this new breeding method is called as " precise breeding ".
The basal core principle of precise breeding is, for often kind of copulation type, as demonstrated in Figure 1, have male (boar and/or its seminal fluid) and female (gilt/sow and/or its ovum) at GN and DN/M place that can be used for producing male and/or Female offspring.In order to use precise breeding generation for the mating plan of copulation type, first subgroup is become based on certain standard by can be used for the male of copulation and/or female classification or " separation ", described standard includes but not limited to, producing the function in pyramid, age, parity, genetic worth (such as, EBV) and/or genetic marker or sudden change.
Table 1.
The percentage ratio (Pij) of the male filial generation in tire (litter) a) produced by male subgroup and female parent's subgroup.
B) based on the age and/or be worth can be used for that these of copulation are male is divided into 5 subgroups.
C) based on the age and/or be worth can be used for that these of copulation are female is divided into 4 subgroups.
In example in Table 1, we have defined for these five male subgroups with for these four female subgroups.A male and jth female sub-generic combinations (each sub-generic combinations represents a copulation hypotype) for each i-th, people can determine the percentage ratio (P of the male filial generation of the maximum increase caused on genetic worth
ij) and the percentage ratio (Q of Female offspring
ij).P
ij+Q
ij＝100。P
ijscope can be 0 to 100.
These are male can be divided into ' a ' individual age subgroup, and is divided into ' b ' individual other subgroup based on its EBV in each age subgroup.Such as, these sows can be divided into ' p ' individual parity subgroup at GN place and in ' q ' at DN/M place individual parity subgroup.In addition, within each parity subgroup, sow can be divided into ' n ' individual other subgroup based on such as its EBV.This will cause for male ' ab ' individual subgroup and ' the individual subgroup of (p+q) n ' produces ' the individual copulation hypotype of ab (p+q) n ' altogether for specific copulation type shown in FIG for female.
In an embodiment of precise breeding, each male and eachly femalely to be treated as a copulation subgroup, and determine the percentage ratio producing the male filial generation of the maximum increase on genetic worth for often kind of possible copulation.
This rule can be applied and have different targets different when.Usually, in order to implement precise breeding technology in a Program of genetic improvement, there are multiple steps that can perform:
A step determines can be used for the exploitation of strain and the resource of propagation.Related resources includes but not limited to, the sow position at GN and DN/M place, the performance test ability at GN and DN place, in the number of hybrid generation of every GN boar (the pure lines boar in the breeding of GN place) commercial farm test and budget.
Another step defines the Program of genetic improvement not using precise breeding technology.In certain embodiments of the present invention, Program of genetic improvement can be designed as and produces for given inbreeding level and/or produce maximum genetic progress with the change of available resources.Staple includes but not limited to, maximum parity, the number being selected for the GN boar of annual breeding, selected GN boar are collected for period (natural law) of breeding, breeding objective and phenotypic data.
Other step is the annual genetic improvement using definitiveness and/or randomization method to estimate about strain exploitation program.
An additional step is the strain exploitation program defining the seminal fluid technology using precise breeding and sex sorting.In certain embodiments of the present invention, except definition strain exploitation program is together with except conventional parameter, this can be used for the male of copulation by comprising and is femalely divided into subgroup.Such as, can ' n ' individual parity subgroup be divided into by female and be divided into ' p ' in each parity subgroup individual EBV subgroup.Individual female subgroup that this causes ' n*p '.Can ' a ' individual age subgroup be divided into by male and be divided into ' b ' in each age subgroup individual EBV subgroup.Individual male subgroup that this causes ' a*b '.Then the matrix exploitation mating plan of ' np ' * ' ab ' copulation hypotype can be used.
Another step is that exploitation is for the mating plan using the strain of precise breeding technology to develop program.Usually, relative to the mating plan (that is, contrast) of strain exploitation program wherein not using precise breeding technology, the inbreeding that the target of such mating plan does not increase increasing genetic progress in this strain.About the mating plan of strain exploitation program using precise breeding technology, can use randomness and/or Deterministic Methods determine relative to contrast what cause in genetic progress, increase (and the increase not in inbreeding) is percentage ratio or the percentage range of male or female filial generation for each copulation hypotype.In certain embodiments of the present invention, can use randomness and/or Deterministic Methods determine relative to contrast what cause the maximum increase in genetic progress (and not increase) in inbreeding is the percentage ratio of male or female filial generation for each copulation hypotype.
An additional step that can perform in the strain exploitation program of use precise breeding technology is the one or more female insemination made with the seminal fluid of sex sorting in a copulation hypotype, to realize the percentage ratio being confirmed as male or female filial generation in a mating plan.For this purpose, can from the one or more male seminal fluid obtaining sex sorting these female identical copulation hypotypes.
In certain embodiments of the present invention, contrast can comprise and any method known in the art can be used to simulate with at those the identical individualities using the strain of precise breeding technology to develop in the mating plan of program to use.In addition, the strain exploitation program using precise breeding technology can be defined in the same manner and develop program about the strain of contrast, except only using except those features found in the strain exploitation program of precise breeding technology.
That put at any time each female only malely can to combinationally use, except non-usage mixed semen with one for using the mating plan of the strain of precise breeding technology exploitation program and contrasting the condition that both can suppose.About contrast, can suppose further, except avoiding the copulation between the individuality that is closely related (as full sib and half sib), be chosen to be female with the available male random mating of parent, and 50/50 being separated of on average having between male and Female offspring of every tire.Alternately, can suppose for using the mating plan of the strain of precise breeding technology exploitation program, the target of male or female scope from 0% to 100% that each copulation hypotype will have for the production of certain average percent, and every tire is substantially to the target of 0%, 50% or 100%.
In certain embodiments of the present invention, the general standard that the mating plan of the strain of precise breeding technology exploitation program only can comprise a framework or derive from the specific objective percentage ratio that each copulation hypotype is determined is used.Such as, general standard can be comprise certain percentage each generation whole male, or young high EBV parent should preferentially produce male.Framework or general standard are implemented as generally increases genetic progress relative to contrast, and does not increase inbreeding.
Another step is included in the copulation in DN/M level in the mating plan with the male of optimized percentage or female pigs, to utilize the seminal fluid of each boar to produce ability best, and utilize superior boar to facilitate genetic improvement and to control inbreeding and the heredity transmission eventually to commercial level.
Alternately or additionally, in certain embodiments of the present invention in order to test or evaluation filial generation in phenotype, genome can be implemented and select or sudden change assisted Selection.For sudden change assisted Selection, GN breeding animals will carry the favourable sudden change of datum object, and some individuality can become extremely important for Program of genetic improvement.Such as, if there are existence three interested sudden changes, and favourable gene frequency is 0.5, then only the allele be found for all three-favours isozygotys by the animal of 1.56%.If the number of the favourable sudden change of qualification increases and/or the frequency of favorable allels reduces, then the percentage ratio with the individuality of ideal genotype will decline.
The application of the best Genetic Contributions theory in precise breeding
Intensity in species (such as pig) to select by the animal in a group between inbreeding and relation increase and add the risk that genetic diversity loses, thus cause being in homozygotic state and being in the deleterious recessive gene of inbreeding decline of higher percentage ratio.Inbreeding is not a problem generally for the existing procedure of breeding, because inbreeding is taken into account by they.Among these existing procedures of breeding, inbreeding is confined to the increase that each generation is less than 1% usually.
The number of full sib that is that produced by restriction at first and/or that select and half sib and by avoiding the copulation between full sib and half sib to control inbreeding.Now, most of procedure of breeding limits inbreeding by the principle implemented based on best Genetic Contributions theory (" OGC ").OGC makes genetic progress maximize, restriction inbreeding rate or the relation between selected candidate are (see Wu Er LeAnn Rimes (Woolliams) J. and Mai Weisen (Meuwissen) T. simultaneously, 1993, " decision rules in breeding program and response difference " (Decision rules andvariance of response in breeding schemes), " animal productiong science " (Anim.Prod.) 56:179-186; And Mai Weisen (Meuwissen) T., 1997, " with predetermined inbreeding rate, Response to selection is maximized " (Maximizing the response of selection with a predefined rate of inbreeding), " animal science magazine " (J.Anim.Sci.) 75:934-940).
The application of OGC in the procedure of breeding generally includes two steps:
1. select the selected parent from selecting candidate, and Genetic Contributions is assigned to from generation to generation next for each selected candidate.
2. exploitation relates to the mating plan of the minimized selected candidate of average inbreeding made in the next one generation.
Be implemented as follows in pig:
Male and the Female offspring of-from generation to generation every or per generation in period (such as weekly) some.
-after performance test, need to carry out selection and determine.Use the generation of OGC theory for the calculating of the best Genetic Contributions of the individuality of each performance test.Rejecting has the individuality of the contribution of the calculating under a certain threshold value (the different threshold values for male and female), and then selected individuality can be used for breeding.
-in each period, many male and femalely can be used for breeding, and algorithm can be used develop and cause the inbred mating plan of minimum average B configuration.
Some commercial packages are had to can be used to implement above concept.For use OGC, available software kit such as GENCONT, EVA (Nordgen) and TGRM
tM(X ' Prime, and for Mating design, if in house software develops infeasible words, purposes (Suo Neisong (Sonesson) A. and Mai Weisen (Meuwissen) T. of simulated annealing can be used, 2000, " mating system that the best contribution with the inbreeding rate of restriction is selected " (Mating schemes for optimum contribution selection with constrained rates ofinbreeding), " selection evolutionary genetics " (Genet.Sel.Evol.) 32:231-248) or X ' Mate
tM(X ' Prime).
In certain embodiments of the present invention, OGC can be implemented limit or control the inbreeding in the strain exploitation program utilizing precise breeding technology.
OGC software needs the structure of pedigree information and the procedure of breeding to illustrate usually, includes but not limited to, the maximum parity of group size, power of test and sow.For use precise breeding technology, also need the definition comprising age and genetic worth subclass.
When using precise breeding technology, replace calculating the best Genetic Contributions for each individuality through performance test, calculate now the best Genetic Contributions of the best Genetic Contributions for the production of each individuality through performance test of male filial generation and each individuality through performance test for the production of Female offspring.Reject and have lower than for the production of male a certain threshold value and the individuality contribute lower than the calculating for the production of female a certain threshold value, and then the individuality selected can be used for breeding to produce male and/or Female offspring.Produce and inbreeding is minimized and the mating plan generating contribution as calculated.
Following table 2 gives the example of the contribution of the calculating of the selection candidate (male and female) producing male and/or Female offspring.
Table 2.
This process can use iteration, most possible evolution algorithmic, to find the best solution meeting all definition and require.In this example, requirement may be:
The maximum level of the inbreeding increase of-definition
The genetic improvement of-given inbreeding restriction maximizes
-each contribution is the multiple of 8.
If the subgroup of definition animal, then the target of each animal is in a group identical.
In production pyramid, the Financial cost be associated with sex sorting process itself is a factor in it is implemented.But, can by using the low dosage insemination techniques that disclose herein and/or relaxing these cost by using the estrus synchrony technology disclosed that every estrous dosage number (insemination) is reduced to one from two herein.
The normally consuming time and costliness of the process of spermatid sample producing sex sorting, generally need to use special flow cytometer apparatus, the technical staff of high discipline and the process of complexity.Regrettably, the typical doses of the boar sperm cell using conventional technology of artificial insemination (as inseminated in cervix uteri) successfully to inseminate required is 1x 10
9individual spermatid is to 3x 10
9individual spermatid, wherein typical boar ejaculum contains about 6x 10
10individual spermatid.Therefore, typical boar ejaculum contains about 20 to 60 artificial insemination doses, this greatly limits the business application of spermatid sample in breeding pig of sex sorting.In addition, as mentioned above, female usually insemination twice in each oestrous cycle.Therefore, if the spermatid number for successfully inseminating can be reduced, then in given time quantum, the artificial insemination dosage of more big figure can be produced for given boar, from commercial standpoint, the utilization for the spermatid sample of sex sorting should be ideal much.In order to the business application in pig of the spermatid sample of broadening sex sorting, some embodiment of the present invention contains low dosage Insemination procedures, comprises via deep intrauterine catheters and laparoscopy insemination and the method making estrus synchrony.These methods make to reduce can using for the option of the number of these male sows of breeding producing in pyramid for the male number of breeding and be thus used for producing in production pyramid of producing.Alternately, replace reducing the male production being used for breeding, by selecting less male for breeding with higher genetic worth in each production rank, the pyramidal heredity of this production can be accelerated through and transmit, finally produce higher-quality market pig.
Only disclose following instance by way of example, these examples not intended to be limits the embodiments of the invention disclosed by any way herein.
Example 1.
In the following example, the average effect using Deterministic Methods prediction to use precise breeding technology in pig breeding program is interested in.
Step 1: definition
do not usethe procedure of breeding (contrast) of precise breeding technology:
Step 1.1: based on the sow group of following variable-definition for the strain at GN place:
The number of-every parity sow
-per period (such as week, year) enters the number of the selected gilt of sow group
-for paternal line with for maternal maximum parity
-age when the production of the first tire and follow-up tire
Step 1.2: the production of definition boar
The number of the boar that-per period (such as week, year) is selected
-for the natural law of the boar of breeding.
-at age of the connatae boar of F1
Step 1.3: filial generation is produced
-available gilt and sow and available boar random mating, and every mold has and is separated with 50/50 between Female offspring male.
-every tire 8 pigletss (4 male and 4 female) can be used for performance test.
-can be used for the number of the male of performance test and Female offspring per period.
Step 1.4: performance test
Eight filial generations tested by-every tire.
-NS
dthe number of the gilt that=per period is selected; NPT
dthe number of=per performance test in period.For selected the part (" p of gilt
d")=NS
d/ NPT
d.Similarly, for selected the part (" p of boar
s")=NS
s/ NPT
s." d " should represent maternal and " s " paternal line should be represented.
-p-value can use the function of p to be converted into selection intensity, or can search in table.
-L
dit is the mean age of the sow (maternal) when offspring born.
-L
sthe mean age of boar (paternal line) when offspring born.
-genetic improvement dG=R
iH* σ
h* (i
d+ i
s)/(L
d+ L
s).Should " σ
h", hereditary variation is a constant, and accuracy R is selected in supposition
iHfor test gilt and boar be identical.Therefore can by comparing (i
d+ i
s)/(L
d+ L
s) value assesses procedure of breeding option.
Step 2: definition
usethe procedure of breeding of precise breeding technology.In addition to the above, following details is important:
Step 2.1: definition sow group:
-based on genetic worth (" sow GGM "), sow is divided into subgroup.
The number of-every " parity * sow GGM " subclass sow.
Step 2.2: definition boar group.
-based on age and genetic worth (" boar GGM "), the boar being used for breeding this period is divided into subgroup.
-for mean age of the boar of each age * boar GGM subgroup when offspring born.
Step 2.3: filial generation is produced
-for each (parity * sow GGM) * (age * boar GGM) combination (that is, for each copulation hypotype) distribute certain percentage have male filial generation to be generated, wherein remainder is Female offspring.Such as, if having two parities and three sow GGM classifications, two boar age categories and two boar GGM classifications, then the percentage ratio of male filial generation needs to be assigned to 24 copulation classifications (that is, 2*3*2*2=24).
-can be used for the number of the male of performance test and Female offspring per period
The male filial generation % that-every parity produces.
The male filial generation % of-every boar age group.
Step 2.4: performance test
-selected gilt and boar are divided into multiple subgroup based on their genetic worth, and this best group of individuals will have higher selection intensity than second-best group etc.Can for each sow GGM and for each boar GGM classification calculating ' i ' value.
Do not having in the procedure of breeding of precise breeding (that is, contrasting situation), these identical sows produce this male and Female offspring.In the procedure of breeding using precise breeding, this male filial generation can be produced with the difference mixing of the sow except this Female offspring.In addition, when contrasting, these identical boars produce this male and Female offspring.Use precise breeding, this male filial generation can be produced with the difference mixing of the boar except this Female offspring.In addition, when using precise breeding, in fact there is the boar of the sow of two types (DD=produces the sow of Female offspring and DS=produces the sow of male filial generation) and two types (boar of SD=generation Female offspring and SS=produce the boar of male filial generation).Therefore, when using precise breeding, genetic improvement dG=R
iH* σ
h* (i
dd+ i
ds+ i
sd+ i
ss)/(L
dd+ L
ds+ L
sd+ L
ss).This σ
hbe a constant, and accuracy R is selected in supposition
iHidentical for the gilt of all tests and boar.Therefore, can by comparing (i
dd+ i
ds+ i
sd+ i
ss)/(L
dd+ L
ds+ L
sd+ L
ss) value assessment procedure of breeding option.
Step 3: use precise breeding that the genetic progress of the procedure of breeding is maximized
Step 3.1: " contrast i/t "=(i
d+ i
s)/(L
d+ L
s).
Step 3.2: the option defined for the percentage ratio of the male filial generation in these tires in often kind of copulation hypotype often plants the Male piglets that copulation hypotype may have 0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% of generation.Such as, assessment will produce (11)
2424 kinds of copulation hypotypes of individual option.
Step 3.3: for each option, " precise breeding i/t "=(i
dd+ i
ds+ i
sd+ i
ss)/(L
dd+ L
ds+ L
sd+ L
ss).
Step 3.4:dG
ratio=" precise breeding i/t " is divided by " contrast i/t ".
Step 3.5: have for dG
ratiothe option of peak close to the best.
Step 3.6: the step with 1% instead of 10% is centered around the option of optimum definition for the percentage ratio of the male filial generation in these tires of each copulation classification of discovery in step 3.5.
Step 3.7: repeat step 3.3 to 3.5 and find optimum.
Alternately, evolution algorithmic can be used to perform step 3.Should ' solution ' start with the solution defined.In our example, in each in 24 kinds of copulation types, this can be every tire 50%/50% male/Female offspring.Then the little deviation of this application definition (such as often kind of copulation type 49/51 and 51/49) and find best solution.Then this program defines little deviation around last solution and finds best new solution etc., until when this solution is stablized.
Step 4: repeat this process for different strategies with different hypothesis
-for paternal or maternal.
-each cycle enters number=50,100,150,200,300 etc. of the gilt of group.
-each cycle is used for number=10,20,30,50,100 etc. of the boar of breeding.
-maximum parity=1,2,3,4,5 or 6.
-for breeding boar cycle=35 days, 70 days, 140 days etc.
-gilt/sow is divided into 2,3,4 ... individual genetic worth classification (or regardless of).
-breeding boar is divided into 2,3,4 ... individual genetic worth classification (or regardless of).
-etc.
Example 2.
The example of the mating plan in the strain exploitation program using precise breeding technology is provided with following table 3 to 5.This mating plan comprises that 30 of being divided into three parity subgroups are female and to be divided into 4 of two male age subgroups male, and each wherein in these male and female subgroups is divided into two genetic worth subgroups further.Assuming that 100% farrowing rate.Table 3 give 24 copulation hypotype classifications each in the number of sow.
Table 3.
A) this table gives every copulation hypotype tire number that (combination of=male subgroup and female subgroup) produces.
B) three subgroups (parity 1,2 and 3) are divided into based on the age by female.
C) two subgroups (1 and 2) are divided into based on the age by male.
D) female in parity is divided into two EBV subgroups (1 and 2) and male in age group is divided into two EBV subgroups (1 and 2).
Female in parity is divided into two EBV subgroups (1 and 2) and male in age group is divided into two EBV subgroups (1 and 2).
For each in 24 kinds of copulation hypotypes, the target for the percentage ratio of the male filial generation of often kind of copulation hypotype is determined relative to contrast, thus the genetic progress in this strain is maximized and inbred increase is remained on the rank of definition.Table 4 gives the target percentage of the male filial generation in each copulation hypotype classification.
Table 4.
A) this table shows the male average percent produced in the tire of every copulation hypotype (combination of=male subgroup and female subgroup).
B) three subgroups (parity 1,2 and 3) are divided into based on the age by female.
C) two subgroups (1 and 2) are divided into based on the age by male.
D) female in parity is divided into two EBV subgroups (1 and 2) and male in age group is divided into two EBV subgroups (1 and 2).
Then can use from table 3 and 4 data determine to produce and can be used for the number of the Male piglets of performance test, as shown in table 5.Under this mating plan, assuming that every tire 8 pigletss can be used for performance test.
Table 5.
A) this table gives every copulation hypotype male number that (combination of=male subgroup and female subgroup) produces.
B) three subgroups (parity 1,2 and 3) are divided into based on the age by female.
C) two subgroups (1 and 2) are divided into based on the age by male.
D) female in parity is divided into two EBV subgroups (1 and 2) and male in age group is divided into two EBV subgroups (1 and 2).
In this mating plan example, can be used for 68 (28%) in all 240 filial generations of performance test for male.
Example 3
Following instance proves, by increasing the male or female number being divided into the standard of subgroup (such as EBV, age, parity, etc.), by increasing the number for the subgroup of given standard, or by the male and female subgroup of definition, people can increase the genetic progress of strain.
The program that contrasts using the program of precise breeding (PB) with the Deterministic Methods used as described in example 1 is compared.
For the program parameter of paternal line
-each cycle 150 the first parity sows
-every 5 months cycles 10 selected boars
-boar is divided into 2 age group and 2 EBV classifications, generates 4 subgroups
-sow is divided into 2 age group (parity 1 and 2) and 3 EBV classifications, generates 6 subgroups
-24 kinds of copulation hypotypes (4*6)
For maternal program parameter
-each cycle 200 the first parity sows
-every 5 months cycles 15 selected boars
-boar is divided into 2 age group and 2 EBV classifications, generates 4 subgroups
-sow is divided into 3 age group (parity 1,2 and 3-5) and 3 EBV classifications, generates 9 subgroups
-36 kinds of copulation hypotypes (4*9).
For the mating plan of strain using precise breeding technology, be malely divided into two age group: " younger male " (using them at first trimester) and " more old male " (follow-up three months periods).These are male is divided into two groups (" HH " and " H ") via their EBV further, and wherein this first group comprises best boar (" HH ") and this second group comprises the boar (" H ") of second the best.The female of paternal line is classified as two age group: " 1 " (parity 1) and " 2 " (parity 2), and maternal these are female is simultaneously divided into three age group: " 1 " (parity 1), " 2 " (parity 2) and " 3 " (parity 3-5).These gilt/sows are divided into three groups (" HH ", " H " and " M ", from being up to minimum breeding value) with equal sizes further based on their EBV.
Contrast: 50% male filial generation in each copulation hypotype
PB-1: the identical optimized percentage of the male filial generation in each copulation hypotype
PB-2: the male optimized percentage in the first tire and in follow-up tire 100% female.
PB-3s: paternal: gilt is used to minimus boar to produce there is the first male tire of optimized percentage.Sow is used to eldest boar to produce there is 100% female second fetus.
PB-3d: maternal: gilt is used to minimus boar to produce to have the first male tire of optimized percentage and sow is used to minimus boar to produce there is 100% female second fetus.Sow is used to eldest boar to produce there is 100% female parity 2-5 tire.
PB-4s: paternal: identical with PB-3s, but utilize now EBV classification.Optimised in each in 6 EBV classifications (2 boar EBV and 3 gilt EBV classifications) of the percentage ratio of male filial generation.
PB-4d: maternal: identical with PB-3d, but utilize now EBV classification.
PB-5: the number increasing EBV classification
PB-6: each individuality is taken as EBV classification and treats.
Assess for program PB-1 to PB-4.Latter two program is have estimated, PB-5 and PB-6 by inferred results.The results are summarized in Fig. 2, show that the complexity level increased in precise breeding causes the larger impact on dG, up to about+10% and maternal middle about+13% in paternal line.The percent of total of male hog drops to about 20% and drop to 15% in maternal in paternal line.
The preparation of the boar sperm cell sample of example 4-sex sorting
The process of the following boar sperm cell sample for the preparation of sex sorting is only provided by way of non-limiting example.The first step in the making of the boar sperm cell sample of sex sorting obtains injection seminal fluid from the boar be applicable to.Once have collected injection seminal fluid, can by its increase-volume in a kind of applicable bulking agent, this bulking agent can comprise a kind of antioxidant.Then can by the part of dilution being rich in sperm of this injection seminal fluid.If this sample needed before sex selection transported, the temperature (typically 16 DEG C-17 DEG C) that then this sample can be maintained at 0 DEG C-39 DEG C continues from about 12 hours to about 18 hours, is transported to flow cytometer for during sex sorting process to it from bleeding point.
Once this spermatid sample is in laboratory, various quality examination can be carried out on this spermatid sample, comprise Survey Operations (such as, via CASA system), vigor (such as, via flow cytometer), form (such as, via microscopy) and concentration (such as, via Nucleo calculating instrument).Then prepare by the spermatid sample of these quality examinations for sorting.
Make this sample by flow cytometer before, with DNA selective dyestuff, this sample is dyeed, be exposed in cancellation formula dyestuff with formed dyeing spermatid sample, be positioned over subsequently in the spermatid source of flow cytometer.Specifically, in certain embodiments, can first with a kind of buffer or bulking agent as BTS (see table 6) by the increase-volume of spermatid sample to can be 100x 10 in some cases
6the final concentration of individual cell/ml, then add DNA selective dyestuff Hoechst 33342 (can in MiliQ water, 5mg/ml; Ref:B-2261), good working concentration can be about 5 μ l/100 1,000,000 cell/ml, but can be used in the DNA dyestuff of the lower and higher concentration within the scope of 0.5 to 20,ul/,100 1,000,000 cell/ml.Then this sample is usually positioned in the sealing bath water between 30 DEG C and 42 DEG C (usually close to 35 DEG C) and continues 10 minutes to 12 hours, except dyeing about 50 minutes, dark area is placed in, then sorting subsequently under room temperature (21 DEG C-22 DEG C).Before this sample of sorting, by this sample filtering to remove large fragment and cell (such as, CellTricks with 0.30 μm), red food dye can be added (when added after filtration, be usually 0.5-5 μ l, or the stock solution 1 μ l of 25mg/ml in MiliQ water) maybe another kind of cancellation formula dyestuff can be added in this sample.Then can use this sample of selected by flow cytometry apoptosis with sheath fluid, this sheath fluid can comprise the component as listed in table 7 in some cases, but also can use other sheath fluids.
Fig. 3 illustrates a part for the flow cytometer forming the spermatid sample of one or more subgroup for sorting with exemplary form, this flow cytometer is represented as 10 usually.In this specific embodiment, carry out sex sorting, like this, these subgroups are the spermatid with X chromosome and the spermatid with Y chromosome.Fig. 3 representative is used for the single technology of sorting sperms, but any known technology for sorting cells known in the art can use together with some embodiment of the present invention.
The flow cytometer 10 of Fig. 3 can be programmed to generate two charged stream of liquid droplets by operator, one containing X-bearing sperm cell, positively charged, 12, a spermatid containing band Y chromosome, electronegative, 13, and uncharged undeflected dead cell stream 14 is only wasted.
Operator can also programme to this flow cytometer by so a kind of way selection, namely, use " high-purity sorting " collection sperm with X chromosome and the sperm with Y chromosome (in other words, the sperm with X chromosome of only living and the sperm with Y chromosome are collected), or use " enrichment sorting " to the programming of this flow cytometer to collect the sperm with X chromosome and the sperm with Y chromosome (in other words, to collect containing the living cells be not previously sorted and by use by use for controlling streaming cell instrument computer can boolean logic gate (Boolean Gate logic) again get rid of the drop of all initial dead cells).Boolean logic gate can also be used collect the only one in the sperm with X chromosome or the sperm with Y chromosome.
At first, spermatid stream under stress deposits to nozzle 15 from spermatid source 11 in such a way, make they can be supplied under the pressure from sheath fluid source 16 sheath fluid in nozzle 15 coaxially around.The agitator 17 that may exist can be very accurately controlled via an agitator controlling organization 18, thus within nozzle 15, produce pressure wave, these pressure waves be passed to this coaxially around spermatid stream (when it leaves nozzle orifice 19).As a result, exit coaxially around spermatid stream 20 finally and regularly can form drop 21.
Make the likely charged of each stream of liquid droplets by cell sensing systems 22, this cell sensing systems comprises one is exited stream 20 illumination laser instrument 23 to nozzle, and the light emission that this fluorescence (fluorescing) flows is detected by sensor 24.The information received by this sensor 24 is fed to separator judgement system 25, and if whether this separator judgement system is with regard to making, molding liquid is charged provides any electric charge to make decision rapidly to this molding drop like this, and then correspondingly makes drop 21 charged.
One of sperm with X chromosome is characterised in that they absorb more fluorescent dye than the sperm with Y chromosome owing to there is more DNA, and Just because of this, be different from the sperm with Y chromosome by the amount at the light launched by this laser excitation with the absorbing dye in the sperm of X chromosome, and this species diversity communicates with separator judgement system 25 with regard to being applied to the charge type of each drop only containing the single spermatid with X chromosome or the spermatid with Y chromosome in theory.Dead cell (or those cells that will be dead) typically absorbs cancellation formula dyestuff, and this communicates as not applying electric charge to the drop containing such cell with separator judgement system 25.
Then these charged or uncharged stream of liquid droplets are passed between a pair static electrification plate 26, make them deflect upward in respective collection container 28 and 29 according to its electric charge one or the other side or not deflect, thus formation have the gender enriched colony of the spermatid with X chromosome of the DNA selective dyestuff be associated with its DNA and the spermatid with Y chromosome of gender enriched accordingly.The subgroup of the uncharged non-deflection containing dead cell (or those cells that will be dead) flows in waste canister 30.
The spermatid sample of this sex sorting be collected in one there is 2.5ml catch in the pipe of the 50ml of liquid, this catches liquid in certain embodiments can be TesTris glucose (TTG) (see table 8) every 2,000 ten thousand cells to 2% egg yolk.In this embodiment, the spermatid sample of this sex sorting will typically have the final volume of about 24ml, about 1x 10
6individual cell/ml.Then this pipe is at room temperature stored about 2 hours in darkroom.
Table 6-BTS bulking agent
| Chemical composition | Sigma's code | G/ liter |
| Glucose | G6152 | 36,941 |
| Sodium citrate | S4641 | 5,999 |
| Sodium bicarbonate | S5761 | 1,261 |
| EDTA | ED2SS | 1,250 |
| Potassium chloride | P3911 | 0,7456 |
| Kanamycin sulfate | K4000 | 0,05 |
Table 7-sheath fluid (PBS)
| Chemical composition | Sigma's code | G/ liter |
| Sodium chloride | S9888 | 8 |
| Potassium chloride | P3911 | 0.2 |
| Biphosphate sodium-hydrate | S9638 | 0.12 |
| Sodium phosphate dibasic heptahydrate | S9390 | 1.717 |
| EDTA | E6758 | 1 |
| Potassium penicillin G | PENK | 0.058 |
| Streptomycin sulfate | S6501 | 0.05 |
Table 8-TesTris glucose (TTG)
| Chemical composition | Sigma's code | g/100ml |
| TES | T1375 | 5 |
| TRIS | T1503 | 0.68 |
| Glucose | G6152 | 0.6 |
| Kanamycin (KANAMICYN) | K4000 | 0.005 |
Once obtain the spermatid sample of sex sorting, it can with the artificial insemination procedures of routine as inseminated in cervix uteri, in vitro fertilization or use together with the artificial insemination of deep intrauterine catheter or laparoscopy.Alternately, for storage, can the spermatid sample of this sex sorting of freezen protective, and then subsequently the time after a while melted for.
The freezen protective of the boar sperm cell sample of example 5-sex sorting
Once make the boar sperm cell sample of sex sorting, optionally by this spermatid sample freezen protective so that transport or storage, can use in the time after a while.Following freezing method can use together with the present invention, but just propose-can use any freezing and storing method known in the art by way of example.
After sizing, the spermatid (having 2,000 ten thousand cells) of the sex sorting contained by the pipe of 50ml can be dispensed in the pipe of 15ml, the wherein sex selection spermatid sample seminal fluid of about 12ml in each pipe, each spermatid containing about 1,000 ten thousand sex sortings.Can be centrifugal 4 minutes at 21 DEG C under 3076g by these pipes.Decant goes out supernatant, and precipitation can retain a little supernatant of about 50 μ l.
Then the first freezing media of the solution that can comprise 20% egg yolk and 80% beta lactose at room temperature can be added to each precipitation.Then the energy of these spermatids can be checked.If can accept, can by these pipe racks to Programmable Temperature controller (PolyScience-MiniTube) upper or can manual handle temperature is reduced to about 5 DEG C from about 21 DEG C through the time of about 2 hours.After fixed time temperature change; these samples can be placed in the cold room of about 5 DEG C; wherein in these samples, add the second freezing media; this second freezing media can comprise egg yolk, beta lactose, glycerol and Equex Stem, or only can comprise cryoprotective agent as glycerol or be cooled to the cryoprotective agent with homeo-osmosis agent of 5 DEG C in advance.After 10 min, can the spermatid sample of sex sorting be placed in artificial insemination suction pipe, then make these suction pipes be exposed in liquid nitrogen vapor (apart from the about 4cm of this liquid nitrogen) and continue short period (such as 10 minutes) and be then placed directly into the medium-term and long-term preservation of liquid nitrogen.
When preparing the semen sample using these sex sortings, thawed by melting/heating (be such as positioned in the water-bath being set in about 37 DEG C and continue about 15 seconds) these suction pipes.Then energy and the vigor of these spermatids can within 30,90 and/or 150 minutes, be analyzed after melting, for standard comparing.
Example 6-estrus synchrony.
For object easily, the present invention considers and can in one or more sow needing to be inseminated, make estrus synchrony and/or bring out timing ovulation.In addition, because the sperm of sex sorting is usually through capacitation in advance, sow insemination within about ovulation 6 hours is importantly made.Estrus synchrony or timing ovulation is made to contribute to guaranteeing that situation is like this.In general, this makes to use one or more hormones or hormone analogs to one or more sow needing to be inseminated.Also exist some induce in gilt rutting period/timing ovulation mode, be described in hereafter.
In order to set up estrus synchrony and ovulation period, one or more hormones or hormone analogs can be used to sow.These hormones and hormone analogs typically comprise, such as, and PG600, OvuGel, eCG, hCG and/or progestogen, and manually can use together with periodical injections, or use by means of a kind of programmable device being placed in sow reproductive tract.The programmable device considered herein discharges one or more hormones or hormone analogs in the mode of time controlled released, and breeder is except programming to the initial parameter for discharging one or more hormones described or hormone analogs, this device need not be monitored or any input is provided.As usually can using together with the present invention for induction and/or the estrous any following method of synchronization of being known in the art of hereafter comprising.
(a)
the rutting period of transport and boar induction.gilt generally reaches puberty when about 180-210 age in days.But, puberty naturally reach the impact being subject to many inherences and extrinsic factor, as genotype, environment and the contact with boar.Many breeders and peasant point out, generally observe rutting period first when gilt is six monthly ages.The again arrangement of estrous beginning usually with animal from gilt multiplier to commercial farm or transport consistent.Certainly, the most famous in pig stressors is transport.If age normally the starting close to puberty of gilt when transporting, then approximately the gilt of 25%-35% will manifest rutting period in one week after shipping.The rutting period of this transport induction can be used for making a part of gilt synchronous.
Although transport can induce rutting period, the contact of boar is the strong form of one stimulated puberty obviously.The principal element controlling the efficiency (stimulating as puberty) of boar contact is the age of the gilt when introducing boar.When starting boar contact at gilt 4 monthly age, puberty reaction is minimum.Someone propose when gilt too young and can not make a response time, childhood gilt may a stage of development to boar stimulate become accustomed to.On the contrary, introduce when boar and be deferred to closest to (6 monthly ages and more than) during first feelings early stage, because this reaction of different reasons is restricted again.Due to the relatively old age (that is, 6 months) of gilt when introducing, actual age puberty of these gilt is a lot of unlike reducing under the animal do not stimulated.When the gilt age interval at 160 days occur boar introduce time, touch the interval of puberty from boar first and be all lowered to bottom line at the gilt age of puberty, the just estrous maximum synchronization of feelings occurs simultaneously.
(b)
oral and time delay discharges progestogen.for this method of estrus synchrony by using the progesterone of oral administration or synthetic progestin and utilizing the suppression of ovarian activity.Some progestogen of time controlled released injectable forms can be obtained, as altrenogest (vide infra).Seriatim or in groups continue 14 to 18 consecutive days with the ratio in the 15-30mg altrenogest/pig/sky gilt be in the cycle of feeding, the last time progestogen feed after synchronously the starting of postpartum estrus between 2 to 8 days.
(c)
promoting sexual gland hormone.eCG/hCG (PG600R) is current, for inducing the most frequently used exogenous hormone combination being in aperiodic female ovarian follicular growth and ovulation to be eCG (being called the priatin (PMSG) of pregnant mare in the past) and the combination of human chorionic gonadotropin (hCG).Product P G600R contains 400IU PMSG and 200IU hCG.This hormone can buy as composition of medicine and for induction be in rutting period of aperiodic pig and ovulation be have cost-benefit.Gilt usually within 3-6 days, demonstrates rutting period after the treatment and ovulation period is about 110-120 hour.If gilt is given boar contact the every day when processing beginning, then this response rate is improved.PG600 comprises priatin (being called horse chorionic-gonadotropin hormone (" PMSG " or " eCG ") in addition) and the human chorionic gonadotropin (" hCG ") (Intervet (Intervet) company) of pregnant mare.OvuGel is the commercially available promoting sexual gland hormone (triptorelin acetate) of the another kind be in controlled-release formulation, it can be used via a kind of Intravaginal delivery system (gel medical science company (Gel Med Sciences, Inc.)).
(d)
prostaglandin.pGF
2α is effective in induction luteolysis, miscarriage and in (and pseudo-fetus) gilt of pregnancy, turned back to rutting period rapidly after conceived more than two week.A kind of is make gilt pen mating (pen-mate) three weeks for synchronized method, then uses PGF after two weeks
2α process.
(e)
time delay releasing hormone.another kind method relates to the commercially available preparation of particular point in time direct injection in the oestrous cycle, as altrenogest or RU-2267.Such as, in one embodiment of the invention, continue over 4 to 7 days, to realize synchronization and timing ovulation by the 15-30mg altrenogest/sky of using for 11-14 days of the oestrous cycle gilt.After stopping altrenogest 24 hours, the PMSG of 400 to 2000IU can be used, after 72 to 83 hours, then use the hCG of 500 to 1000IU.
Example 7-ovulation tests.
Can by checking that the follicle of sow completes the ovulation tests in sow.Critical in the test-tube management of the realization of setting up the importance of sufficient sperm bank at the appropriate time relative to ovulation in fallopian tube in pig.Specifically, when knowing of probably ovulating during rutting period of sow is very beneficial for realizing successfully insemination.For this reason, in a particular embodiment of the present invention, after inducing rutting period, use the follicle of ultrasonic examination sow.In one particular embodiment of the present invention, 30 hours after hCG injection start, by the ovary that there is an every 4 hour inspection sow of Transrectal Ultrasound for preovulatory follicle.The sow selecting the multiple preovulatory follicle of display (follicular antrum diameter >6mm) for 2-3 hour after ultrasonic examination is used for insemination.
Example 8-uses laparoscopy or the insemination of deep intrauterine catheter
Once prepare the boar semen sample of sex sorting, this sample can be used for sow is inseminated.The technology of artificial insemination of any routine can be used in the present invention, comprise insemination in cervix uteri.But, deep intrauterine catheter and laparoscopy are significant especially in pig, because they allow to use the spermatid reducing dosage to be used for successful fertilization, partly because spermatid can be placed in the key area of the reproductive tract of sow by they, include but not limited to, cornua uteri, fallopian tube, ampulla, isthmus and uterotubal junction.Use the spermatid dosage reduced to allow the boar superior in heredity using much less to be used for breeding objective, this has the cost reducing breeder and the benefit reduced owing to must support the environmental hazard caused by a large amount of boar.
A () uses deep intrauterine catheter insemination.Deep intrauterine catheter is used to allow people to be inserted by spermatid in the cornua uteri of sow and be in uterotubal junction ideally.A kind of like this deep intrauterine catheter use and structure is disclosed in U.S. Patent number 6,695, in 767, its disclosure content combines in full with it hereby by reference.So a kind of deep intrauterine catheter optionally can comprise a video camera or endoscope, with the path allowing operator to see this conduit, still can make one's options between two cornua uteris like this being placed on by spermatid one.Alternately, when being combined with radiography device or fluoroscopic equipment, this deep intrauterine catheter position within sow reproductive tract can be visualized.Due to its length, deep intrauterine catheter allows operator to reach the remote area of the reproductive tract of sow, comprises that cornua uteri-this uses for the unreachable region of test-tube standard catheter.In one embodiment of the invention, the length of this deep intrauterine catheter is 1.8m, 1-2m, 1-2.5m, 1-3m, 2-3m, 2-3.5m or 2.5-3m.
This deep intrauterine catheter can be introduced into the inner side of the cervical canal being in sow in rutting period, and this sow can be superovulated, but also can be free period or otherwise induce.Can by non-toxic aq lubricant applications to this conduit so that it is through vagina.This conduit can comprise an outer tube or sheath and a flexible probe within this outer tube or sheath.In one embodiment of the invention, once this conduit advances to cervical canal, then this flexible probe can be advanced further within the outer tube of this conduit.This flexible probe can be advanced until reach the previous section of cornua uteri.When this flexible probe is advanced in cornua uteri, it can bend and the zigzag path therefore continued along cornua uteri is advanced.Although be not imperative, introduce liquid in a small amount by the outer tube of this conduit and this flexible probe can be promoted to advance in its passage by cervical canal and advance through cornua uteri.Once this flexible probe has been introduced into the final position reached in cornua uteri, the spermatid sample be included in a syringe can be connected to the near-end of this flexible probe, and can be incorporated into by the flexible duct in this flexible probe in the environment of uterus.In order to avoid spermatid loss and guarantee that this spermatid sample is complete emptying from this flexible duct, subsequently can by this flexible duct introduce gobbet.After this, the conduit comprising this outer tube and this flexible probe can be recalled.In another aspect of the present invention, this process also may be used for embryo to be delivered in cornua uteri or from cornua uteri to shift out embryo.
B () uses laparoscopy insemination.The advantage using laparoscopy that sow is inseminated is, the placement of spermatid in the reproductive tract of sow can be even more more accurate than use conduit, therefore makes it possible to further use the spermatid dosage reduced for insemination.Can the uterotropic specific region of target, as fallopian tube, isthmus, ampulla or uterotubal junction.As limiting examples, follow procedure can use sow and inseminate via laparoscopy together with the present invention.
Such as, about 1x 10 can be had by what contain in 50ml pipe
6the spermatid sample of the sex sorting of the 24ml of individual spermatid/ml to be dispensed in the pipe of 2 15ml and in about 3076g temperature centrifugal a few minutes (2-5 or 4 minute) within the scope of about 21 DEG C.If needed, can by supernatant recentrifuge under the same conditions.Then the seminal fluid of generation precipitation is mixed and checks concentration (via Nucleo calculating instrument).With BTS by the spermatid diluted sample of concentrated sex sorting to 10x 10
6the final concentration of individual cell/ml also checks energy and the vigor of these spermatids.(under in this whole process, this spermatid sample should be maintained at room temperature (21 DEG C).)
By sow grouping or can separate, such as, they can be assigned to separately in the force ventilation restriction facility of matting.Sow (2 – 6 parity) was wean in the about the 21st day.Then latter 24 hours can by the horse chorionic-gonadotropin hormone (eCG with about 1250IU in wean; Folligon, Intervet Internat B. V. (Intervet InternationalB.V.), Bock Si Meier (Boxmeer), Holland-or a kind of equivalent compound) intramuscular injection is each femalely induces rutting period; After 72 hours, with about 750IU human chorionic gonadotropin (hCG; VeterinCorion, Di Washa (Divasa), Farmavic S.A., Barcelona, Spain) or a kind of equivalent they are processed.Carry out monoestrus phase detection (such as at 7:00 in the morning) every day, 2 days after eCG injection start.Detecting estrous a kind of mode is allow female and ripe boar nose to contact nose, and identifies by applying back-pressure the sow showing quiet vertical reflection of oestrusing, and it is considered and is in rutting period; Can scan ovary at this moment.Use many scanning angles transducer of 5MHz by transrectal ultrasonography, the mature follicle checking ovary with period distances (such as every 4 hours) can be started after hCG injects about 30 hours, to find the existence of preovulatory follicle.The sow of the multiple preovulatory follicle of display (follicular antrum diameter >6mm) is only had just to be selected for insemination.After ultrasonic examination, 2 – inseminated within 3 hours.
Then, once these sows (can be used by means of azaperone by calmness; Grant Jingning (Stresnil); 2mg/kg body weight, intramuscular injection), peritoneoscope insemination can be carried out on these sows.Also can with a kind of compound as penthiobarbital (Abbott (Abbot); 7mg/kg body weight, intravenous injection) induce as general anesthesia and maintain with halothane (3.5% – 5%) or a kind of similar compound.For operation, sow can be placed in dorsal position, and if available, make it lie on the back in celioscope bracket.If use support, to be placed on Te Lundelunbai (Trendelenburg) clinostatism (hind leg upwards, under head points) higher than the angle of about 20 ° of horizontal plane.
In one embodiment, an otch (about 1.5cm) is being done close to umbilical part.Then with countertraction upwards this otch of tractive edge and make the Optiview trocar (KMS Medical LLC (Ethicon Endo-surgery) of 0 ° of laparoscopically 12mm with insertion, Cincinnati, Ohio (Cincinnati OH), USA) advance in this wound.At umbilicus place, by means of slight cutting and modest pressure, through fascia, transverse fascia and peritoneum after fascia, rectus, rectus before subcutaneus adipose tissue, rectus.This process is controlled via supervision feedback.Although CO
2pipe is connected on this trocar, but until just starts inflation when this peritoneum is pierced.After entering peritoneal cavity and starting to form pneumoperitoneum, remove the handpiece of Optiview, and replace with 0 ° of peritoneoscope.Use CO
2abdominal cavity is inflated to 14mmHg.Two accessory aperture are placed in half abdominal part of right side and left part, and its peritoneoscope Duval each provided for handling cornua uteri clamps and grasps oviducal entrance for pin of inseminating.Fallopian tube is grasped with the Duval pincers in isthmus region.Then dosage stream (containing 30-50 ten thousand sperms in 0.1ml) is inserted, and the sperm of sex sorting is flushed in fallopian tube.Then in another fallopian tube, this program is repeated.After two oviductus lateraliss are all by insemination, the trocar removed and sews up these incisional wounds.
Persons of ordinary skill in the art will recognize that the invention described above comprises many inventive embodiments, at least comprise the following:
A. increase a method for the genetic worth of pig, the method comprises the following steps:
Set up the multiple copulation hypotypes for a strain;
To determine for each in these copulation hypotypes be the percentage ratio of male filial generation or be the percentage ratio of female filial generation for each in these copulation hypotypes, and this percentage ratio will cause genetic worth increase in this strain relative to contrast;
Divided by spermatid sample from the male hog be in one of these copulation hypotypes and elect one or more spermatid subgroup as, the spermatid of in one of them spermatid subgroup at least 60% is with X chromosome or Y chromosome; And
The one or more female pigs inseminations be in one of these copulation hypotypes are made, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast with the spermatid of this subgroup.
A1. the method as described in A, is wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
A2. the method as described in A or A1, wherein this strain comprises a paternal line or maternal.
A3. the method according to any one of A to A2, wherein in this contrast, the filial generation of about 50% is male.
A4. the method according to any one of A to A3, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
A5. the method according to any one of A to A4, the classification of the male hog wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or age.
A6. the method according to any one of A to A5, the classification of the female pigs wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or parity.
A7. the method according to any one of A to A6, the percentage ratio of the spermatid wherein in the spermatid subgroup with X chromosome or Y chromosome is selected from lower group, and this group is made up of the following: at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and at least 100%.
A8. the method according to any one of A to A7, wherein this strain comprises a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35%.
A9. the method as described in A8 is wherein that the percentage ratio of male filial generation is between about 15% to 30% for these product.
A10. the method according to any one of A to A7, wherein this strain comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30%.
A11. the method as described in A10 is wherein that the percentage ratio of male filial generation is between about 10% to 25% for these product.
A12. the method according to any one of A to A7, wherein for each in these copulation hypotypes be male filial generation percentage ratio or for each in these copulation hypotypes be the percentage ratio of female filial generation between about 0 to 10% or between about 90% to 100%, and the filial generation of any generation is the member of daughter nucleus or multiplier.
A13. the method according to any one of A to A12, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain or an only female pigs.
A14. the method according to any one of A to A13, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain and an only female pigs.
A15. the method according to any one of A to A14, wherein these percentage ratios determined use randomness or Deterministic Methods to determine.
A16. the method according to any one of A to A15, wherein male or female genetic worth is the function of one or more maternal character.
A17. the method according to any one of A to A15, wherein male or female genetic worth is the function of one or more paternal character.
A18. the method according to any one of A or A15, wherein male or female genetic worth is the function of selection index.
A19. the method according to any one of A or A15, wherein male or female genetic worth is the function of EBV.
A20. the method as described in A18, wherein this selection index is the function deriving from the data comprising this male or this female group.
A21. the method as described in A16, wherein these character comprise fertility, tire litter size and milk yield.
A22. the method as described in A17, wherein these character comprise feed efficiency, average daily gain and carcass lean meat percentage.
A23. the method according to any one of A to A22, the deep intrauterine catheter wherein making one or more female pigs inseminate to comprise and use and inserted by the film of one or more female pigs or pin use this subgroup to the reproductive tract of this one or more female pigs.
A24. the method as described in A23, wherein uses deep intrauterine catheter to use this subgroup to the reproductive tract of one or more female pigs and comprises and being inserted in one or more cornua uteri by described spermatid.
A25. the method as described in A23 or A24, wherein said deep intrauterine catheter comprises a video camera or endoscope.
A26. the method according to any one of A23 to A25, comprises the step made via radiography or fluoroscopy in the visual reproductive tract simultaneously inserting described sow of this deep intrauterine catheter further.
A27. the method according to any one of A to A26, wherein this subgroup comprises 1x 109 or less spermatid.
A28. the method as described in A23, is wherein used the pin that inserted by the film of this one or more female pigs to use this subgroup to the reproductive tract of this one or more female pigs and comprises and this subgroup being expelled in one or more fallopian tube of this one or more female pigs.
A29. the method as described in A28, comprises further and makes via a peritoneoscope or video camera the visual step of described pin that is inserted in described one or more fallopian tube.
A30. the method according to any one of A to A23 and A28 to A29, wherein this subgroup comprises 1x 10
6individual or less spermatid.
A31. the method according to any one of A to A30, comprises the step making Estrus synchronization in this one or more female pigs or induction timing ovulation by using one or more hormones or hormone analogs to one or more female pigs further.
A32. the method as described in A31, wherein these one or more hormones or hormone analogs comprise PG600, OvuGel, eCG, progestogen, hCG, altrenogest or RU-2267.
A33. the method as described in A31 or A32, wherein these one or more hormones or hormone analogs are that programmable device in reproductive tract by being positioned over this one or more female pigs is used.
A34. the method according to any one of A to A33, comprises further by checking that one or more follicles of this one or more female pigs detect the step of the ovulation in this one or more female pigs.
A35. the method as described in A34, wherein this one or more follicle is that use is ultrasonographic.
A36. the method according to any one of A to A35, comprises the other step carrying out Selection parent based on phenotype measurement.
A37. the method according to any one of A to A35, comprises the other step selecting the parent for this strain based on genotype.
A38. the method according to any one of A to A35, comprises the step that other use sudden change assisted Selection carrys out Selection parent.
A39. the method according to any one of A to A35, wherein genetic worth measures based on phenotype.
A40. the method according to any one of A to A35, wherein genetic worth is based on genotype.
A41. the method according to any one of A to A40, wherein this male hog or this one or more female pigs are the members of hereditary core, daughter nucleus or multiplier.
A42. the method according to any one of A to A40, wherein this male hog or this one or more female pigs are the members of hereditary core.
A43. the method according to any one of A to A42, the step wherein set up for multiple copulation hypotypes of a strain performs as the part of a procedure of breeding.
A44. the method according to any one of A to A42, the step wherein set up for multiple copulation hypotypes of a strain performs as generation part for the mating plan of this strain.
A45. the method according to any one of A to A44, is wherein made up of a male subgroup or a female subgroup for each in these copulation hypotypes of this strain.
A46. the method as described in A45, wherein this male subgroup or this female subgroup are defined by one or more standard or based on one or more standard, and described standard is included in the function of producing in pyramid, parity, age, genetic worth, genetic marker or genetic mutation.
B. increase a method for the genetic worth of pig, the method comprises the following steps:
Set up the multiple copulation hypotypes for a strain; And
Determine to be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what cause in this strain genetic worth to increase or to be the percentage ratio of female filial generation for each in these copulation hypotypes.
B1. the method as described in B, is wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
B2. the method as described in B or B1, wherein this strain comprises a paternal line or maternal.
B3. the method according to any one of B to B2, wherein in this contrast, the filial generation of about 50% is male.
B4. the method according to any one of B to B3, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
B5. the method according to any one of B to B4, the classification of the male hog wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or age.
B6. the method according to any one of B to B5, the classification of the female pigs wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or parity.
B7. the method according to any one of B to B6, wherein in this first subgroup at least 80% spermatid with X chromosome or wherein in this first subgroup the spermatid of at least 80% with Y chromosome.
B8. the method according to any one of B to B7, wherein this strain comprises a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35%.
B9. the method as described in B8 is wherein that the percentage ratio of male filial generation is between about 15% to 30% for these product.
B10. the method according to any one of B to B7, wherein this strain comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30%.
B11. the method as described in B10 is wherein that the percentage ratio of male filial generation is between about 10% to 25% for these product.
B12. the method according to any one of B to B7, wherein for each in these copulation hypotypes be male filial generation percentage ratio or for each in these copulation hypotypes be the percentage ratio of female filial generation between about 0 to 10% or between about 90% to 100%, and the filial generation produced is the member of daughter nucleus or multiplier.
B13. the method according to any one of B to B12, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain or an only female pigs.
B14. the method according to any one of B to B13, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain and an only female pigs.
B15. the method according to any one of B to B14, wherein these percentage ratios determined use randomness or Deterministic Methods to determine.
B16. the method according to any one of B to B15, wherein male or female genetic worth is the function of one or more maternal character.
B17. the method according to any one of B to B15, wherein male or female genetic worth is the function of one or more paternal character.
B18. the method according to any one of B or B15, wherein male or female genetic worth is the function of selection index.
B19. the method according to any one of B or B15, wherein male or female genetic worth is the function of EBV.
B20. the method as described in B18, wherein this selection index is the function deriving from the data comprising this male or this female group.
B21. the method as described in B16, wherein these character comprise fertility, tire litter size and milk yield.
B22. the method as described in B17, wherein these character comprise feed efficiency, average daily gain and carcass lean meat percentage.
C. be increased in a method for the genetic worth in paternal line, the method comprises the following steps:
Divided by one or more spermatid samples from the one or more male hogs in this paternal line and elect one or more spermatid subgroup as, in one of them subgroup, the spermatid of at least 70% is with X chromosome or Y chromosome; And
The one or more female pigs insemination in this paternal line is made, to realize being the percentage ratio of male filial generation for this paternal line between about 10% to 35% by this subgroup.
C1. the method as described in C is wherein that the percentage ratio of male filial generation is between about 15% to 30% for this paternal line.
C2. the method as described in C or C1, the deep intrauterine catheter wherein making one or more female pigs inseminate to comprise and use and inserted by the film of one or more female pigs or pin use this subgroup to the reproductive tract of this one or more female pigs.
C3. the method as described in C2, wherein uses deep intrauterine catheter to use this subgroup to the reproductive tract of one or more female pigs and comprises and being inserted in one or more cornua uteri by described spermatid.
C4. the method as described in C2 or C3, wherein said deep intrauterine catheter comprises a video camera or endoscope.
C5. the method according to any one of C2 to C4, comprises the step made via radiography or fluoroscopy in the visual reproductive tract simultaneously inserting described sow of this deep intrauterine catheter further.
C6. the method according to any one of C to C5, wherein this subgroup comprises 1x 10
9individual or less spermatid.
C7. the method as described in C2 or C6, is wherein used the pin that inserted by the film of this one or more female pigs to use this subgroup to the reproductive tract of this one or more female pigs and comprises and this subgroup being expelled in one or more fallopian tube of this one or more female pigs.
C8. the method as described in C7, comprises further and makes via a peritoneoscope or video camera the visual step of described pin that is inserted in described one or more fallopian tube.
C9. the method according to any one of C to C8, wherein this subgroup comprises 1x 10
6individual or less spermatid.
C10. the method according to any one of C to C9, comprises the step making Estrus synchronization in this one or more female pigs or induction timing ovulation by using one or more hormones or hormone analogs to one or more female pigs further.
C11. the method as described in C10, wherein these one or more hormones or hormone analogs comprise PG600, OvuGel, eCG, progestogen, hCG, altrenogest or RU-2267.
C12. the method as described in C10 or C11, wherein these one or more hormones or hormone analogs are that programmable device in reproductive tract by being positioned over this one or more female pigs is used.
C13. the method according to any one of C to C12, comprises further by checking that one or more follicles of this one or more female pigs detect the step of the ovulation in this one or more female pigs.
C14. the method as described in C13, wherein this one or more follicle is that use is ultrasonographic.
D. be increased in maternal in the method for genetic worth, the method comprises the following steps:
Divided by one or more spermatid samples of the one or more male hogs in maternal from this and elect one or more spermatid subgroup as, in one of them subgroup, the spermatid of at least 70% is with X chromosome or Y chromosome; And
One or more female pigs insemination in making this maternal by this subgroup, with realize between about 5% to 30% for this maternal percentage ratio for male filial generation.
D1. the method as described in D, wherein maternal for this is that the percentage ratio of male filial generation is between about 10% to 25%.
D2. the method as described in D or D1, the deep intrauterine catheter wherein making one or more female pigs inseminate to comprise and use and inserted by the film of one or more female pigs or pin use this subgroup to the reproductive tract of this one or more female pigs.
D3. the method as described in D2, wherein uses deep intrauterine catheter to use this subgroup to the reproductive tract of one or more female pigs and comprises and being inserted in one or more cornua uteri by described spermatid.
D4. the method as described in D2 or D3, wherein said deep intrauterine catheter comprises a video camera or endoscope.
D5. the method according to any one of D2 to D4, comprises the step made via radiography or fluoroscopy in the visual reproductive tract simultaneously inserting described sow of this deep intrauterine catheter further.
D6. the method according to any one of D to D5, wherein this subgroup comprises 1x 10
9individual or less spermatid.
D7. the method as described in D2 or D6, is wherein used the pin that inserted by the film of this one or more female pigs to use this subgroup to the reproductive tract of this one or more female pigs and comprises and this subgroup being expelled in one or more fallopian tube of this one or more female pigs.
D8. the method as described in D7, comprises further and makes via a peritoneoscope or video camera the visual step of described pin that is inserted in described one or more fallopian tube.
D9. the method according to any one of D to D8, wherein this subgroup comprises 1x 10
6individual or less spermatid.
D10. the method according to any one of D to D9, comprises the step making Estrus synchronization in this one or more female pigs or induction timing ovulation by using one or more hormones or hormone analogs to one or more female pigs further.
D11. the method as described in D10, wherein these one or more hormones or hormone analogs comprise PG600, OvuGel, eCG, progestogen, hCG, altrenogest or RU-2267.
D12. the method as described in D10 or D11, wherein these one or more hormones or hormone analogs are that programmable device in reproductive tract by being positioned over this one or more female pigs is used.
D13. the method according to any one of D to D12, comprises further by checking that one or more follicles of this one or more female pigs detect the step of the ovulation in this one or more female pigs.
D14. the method as described in D13, wherein this one or more follicle is that use is ultrasonographic.
E. one kind increases the method for the genetic worth of pig, the method comprises the multiple copulation hypotype by setting up for strain and determines for each percentage ratio for male filial generation in these copulation hypotypes or the mating plan each percentage ratio for female filial generation in these copulation hypotypes being produced to a strain for pig, the increase on genetic worth that these filial generations will cause relative to contrast.
E1. the method as described in E, is wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
E2. the method as described in E or E1, wherein this strain comprises a paternal line or maternal.
E3. the method according to any one of E to E2, wherein in this contrast, the filial generation of about 50% is male.
E4. the method according to any one of E to E3, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
E5. the method according to any one of E to E4, the classification of the male hog wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or age.
E6. the method according to any one of E to E5, the classification of the female pigs wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or parity.
E7. the method according to any one of E to E6, wherein in this first subgroup at least 80% spermatid with X chromosome or wherein in this first subgroup the spermatid of at least 80% with Y chromosome.
E8. the method according to any one of E to E7, wherein this strain comprises a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35%.
E9. the method as described in E8 is wherein that the percentage ratio of male filial generation is between about 15% to 30% for these product.
E10. the method according to any one of E to E7, wherein this strain comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30%.
E11. the method as described in E10 is wherein that the percentage ratio of male filial generation is between about 10% to 25% for these product.
E12. the method according to any one of E to E7, wherein for each in these copulation hypotypes be male filial generation percentage ratio or for each in these copulation hypotypes be the percentage ratio of female filial generation between about 0 to 10% or between about 90% to 100%, and the filial generation produced is the member of daughter nucleus or multiplier.
E13. the method according to any one of E to E12, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain or an only female pigs.
E14. the method according to any one of E to E13, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain and an only female pigs.
E15. the method according to any one of E to E14, wherein these percentage ratios determined use randomness or Deterministic Methods to determine.
E16. the method according to any one of E to E15, wherein male or female genetic worth is the function of one or more maternal character.
E17. the method according to any one of E to E15, wherein male or female genetic worth is the function of one or more paternal character.
E18. the method according to any one of E or E15, wherein male or female genetic worth is the function of selection index.
E19. the method according to any one of E or E15, wherein male or female genetic worth is the function of EBV.
E20. the method as described in E18, wherein this selection index is the function deriving from the data comprising this male or this female group.
E21. the method as described in E16, wherein these character comprise fertility, tire litter size and milk yield.
E22. the method as described in E17, wherein these character comprise feed efficiency, average daily gain and carcass lean meat percentage.
E23. the method according to any one of E to E22, wherein this strain belongs to hereditary core, daughter nucleus or multiplier.
E24. the method according to any one of E to E22, wherein this strain belongs to hereditary core.
E25. the method according to any one of E to E24, the step wherein set up for multiple copulation hypotypes of a strain performs as the part of a procedure of breeding.
E26. the method according to any one of E to E24, the step wherein set up for multiple copulation hypotypes of a strain performs as generation part for the mating plan of this strain.
F. increase a method for the genetic worth of pig, the method comprises the following steps:
Set up the multiple copulation hypotypes for a strain;
To determine for each in these copulation hypotypes be the percentage ratio of male filial generation or be the percentage ratio of female filial generation for each in these copulation hypotypes, and this percentage ratio will cause genetic worth increase in this strain relative to contrast;
Divided by spermatid sample from the male hog be in one of these copulation hypotypes and elect one or more spermatid subgroup as, the spermatid of in one of them spermatid subgroup at least 60% is with X chromosome or Y chromosome; And
The one or more ovum fertilizations from the one or more female pigs be in one of these copulation hypotypes are made, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast with the spermatid of this subgroup.
F1. the method as described in F, the step of wherein this fertilization completes in vivo.
F2. the method as described in F, the step of wherein this fertilization completes in vitro.
F3. the method according to any one of F to F2, is wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
F4. the method according to any one of F to F3, wherein this strain comprises a paternal line or maternal.
F5. the method according to any one of F to F4, wherein in this contrast, the filial generation of about 50% is male.
F6. the method according to any one of F to F5, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
F7. the method according to any one of F to F6, the classification of the male hog wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or age.
F8. the method according to any one of F to F7, the classification of the female pigs wherein in one or more copulation hypotype defines by the one or more features comprising genetic worth or parity.
F9. the method according to any one of F to F8, the percentage ratio of the spermatid wherein in the spermatid subgroup with X chromosome or Y chromosome is selected from lower group, and this group is made up of the following: at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and at least 100%.
F10. the method according to any one of F to F9, wherein this strain comprises a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35%.
F11. the method as described in F10 is wherein that the percentage ratio of male filial generation is between about 15% to 30% for these product.
F12. the method according to any one of F to F9, wherein this strain comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30%.
F13. the method as described in F12 is wherein that the percentage ratio of male filial generation is between about 10% to 25% for these product.
F14. the method according to any one of F to F9, wherein for each in these copulation hypotypes be male filial generation percentage ratio or for each in these copulation hypotypes be the percentage ratio of female filial generation between about 0 to 10% or between about 90% to 100%, and the filial generation produced is the member of daughter nucleus or multiplier.
F15. the method according to any one of F to F14, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain or an only female pigs.
F16. the method according to any one of F to F15, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain and an only female pigs.
F17. the method according to any one of F to F16, wherein these percentage ratios determined use randomness or Deterministic Methods to determine.
F18. the method according to any one of F to F17, wherein male or female genetic worth is the function of one or more maternal character.
F19. the method according to any one of F to F17, wherein male or female genetic worth is the function of one or more paternal character.
F20. the method according to any one of F to F17, wherein male or female genetic worth is the function of selection index.
F21. the method according to any one of F to F17, wherein male or female genetic worth is the function of EBV.
F22. the method as described in F20, wherein this selection index is the function deriving from the data comprising this male or this female group.
F23. the method as described in F18, wherein these character comprise fertility, tire litter size and milk yield.
F24. the method as described in F19, wherein these character comprise feed efficiency, average daily gain and carcass lean meat percentage.
F25. the method according to any one of F to F24, comprises the other step carrying out Selection parent based on phenotype measurement.
F26. the method according to any one of F to F24, comprises the other step selecting the parent for this strain based on genotype.
F27. the method according to any one of F to F24, comprises the step that other use sudden change assisted Selection carrys out Selection parent.
F28. the method according to any one of F to F27, wherein this male hog or this one or more female pigs are the members of hereditary core, daughter nucleus or multiplier.
F29. the method according to any one of F to F27, wherein this male hog or this one or more female pigs are the members of hereditary core.
F30. the method according to any one of F to F29, the step wherein set up for multiple copulation hypotypes of a strain performs as the part of a procedure of breeding.
F31. the method according to any one of F to F29, the step wherein set up for multiple copulation hypotypes of a strain performs as generation part for the mating plan of this strain.
G. increase a method for the strain of pig or the genetic progress of kind, the method comprises the following steps:
Semen sample is collected from the boar from described strain or kind;
Divided by described semen sample and elect at least two spermatid subgroups as, wherein first subgroup of at least 80% is with X chromosome or Y chromosome;
The sow insemination from described strain or kind is made with the spermatid from described first subgroup;
Offspring is produced from described sow; And
Calculate the selection index for one or more described offspring;
Select the one or more described offspring compared to the average selection index for described strain or kind with higher selection index, to carry out breeding with the pig from described strain or kind, thus increase the genetic progress of described strain or kind.
G1. the method as described in G, wherein said strain or kind are gilt strains, and described first subgroup is with X chromosome.
G2. the method as described in G, wherein said strain or kind are boar strains, and described first subgroup is with Y chromosome.
G3. the method as described in G, the selection index wherein for one or more described offspring carries out calculating based on the data deriving from the group comprising described offspring.
G4. the method as described in G, the described selection index wherein for one or more described offspring comprises measurement fertility, tire litter size and milk yield character.
G5. the method as described in G, the described selection index wherein for one or more described offspring comprises measurement feed efficiency, average daily gain and carcass lean meat percentage character.
G6. the method as described in G, the deep the intrauterine catheter wherein step from the sow insemination of described strain or kind being comprised use and inserted by the film of described sow with the spermatid from described first subgroup or pin use described spermatid to the reproductive tract of described sow.
G7. the method as described in G6, wherein uses deep intrauterine catheter to use described spermatid to the reproductive tract of described sow and comprises and being inserted in one or more cornua uteri by described spermatid.
G8. the method as described in G7, wherein said deep intrauterine catheter comprises a video camera or endoscope.
G9. the method as described in G7, comprises the step made via radiography or fluoroscopy in the visual reproductive tract simultaneously inserting described sow of this deep intrauterine catheter further.
G10. the method as described in G7, the sum of wherein used spermatid is 1x 10
9individual or less spermatid.
G11. the method as described in G6, is wherein used the pin that inserted by the film of described sow to use described spermatid to the reproductive tract of described sow and comprises described spermatid injection in one or more fallopian tube in the uterus of described sow.
G12. the method as described in G11, comprises further and makes via a peritoneoscope or video camera the visual step of described pin that is inserted in described one or more fallopian tube.
G13. the method as described in G11, the sum of wherein used spermatid is 1x 10
6individual or less spermatid.
G14. the method according to any one of G to G13, comprises the step making Estrus synchronization in described sow or induction timing ovulation by using one or more hormones or hormone analogs to described sow further.
G15. the method as described in G14, one or more hormones wherein said or hormone analogs comprise PG600, OvuGel, eCG, progestogen or hCG.
G16. the method as described in G14, one or more hormones wherein said or hormone analogs are that the programmable device in the reproductive tract by being positioned over described sow is used.
G17. the method as described in G14, comprises further by checking that described female follicle detects the step of the ovulation in described sow.
G18. the method as described in G17, wherein said follicle is that use is ultrasonographic.
G19. the method according to any one of G to G13, wherein said sow is the member of hereditary core or multiplier group.
G20. the method according to any one of G to G13, wherein said boar is the member of hereditary core or multiplier group.
H. increase a method for the strain of pig or the genetic progress of kind, the method comprises the following steps:
Semen sample is collected from the boar from described strain or kind;
Divided by described semen sample and elect at least two spermatid subgroups as, wherein first subgroup of at least 80% is with X chromosome or Y chromosome;
The sow insemination from described strain or kind is made with the spermatid from described first subgroup;
Offspring is produced from described sow;
Obtain the value for the character in one or more described offspring; And
Select one or more described offsprings with the value for described character of the meansigma methods be greater than or less than for the described character in described strain or kind, to carry out breeding with the pig from described strain or kind, thus to increase the genetic progress of described strain or kind.
H1. the method as described in H, wherein said strain or kind are gilt strains, and described first subgroup is with X chromosome.
H2. the method as described in H, wherein said strain or kind are boar strains, and described first subgroup is with Y chromosome.
I. be increased in a method for the number of the offspring of the boar superior in heredity in swinery or on pig farm, the method comprises:
Set up the subgroup from one or more boars superior in heredity of the boar group on a group or a farm;
Spermatid sample is obtained from one or more boar superior on heredity;
Multiple spermatid is prepared from each these spermatid samples;
Multiple sows in described group or on described farm use one or more hormones or hormone analogs, to each sow induction timing ovulation; And
Use deep intrauterine catheter or laparoscopic surgery one or more spermatid dosage that sow is inseminated, this one or more spermatid dosage being wherein administered to each sow altogether comprises and is less than 1x 10
9individual spermatid sum;
Be increased in the number of the offspring of the boar superior in heredity in swinery or on pig farm thus.
J. reduce in swinery or on pig farm for a method for the number of the required boar of breeding, the method comprises:
Set up the subgroup from one or more boars superior in heredity of the boar group on a group or a farm;
Spermatid sample is obtained from one or more boar superior on heredity;
Multiple spermatid is prepared from each these spermatid samples;
Multiple sows in described group or on described farm use one or more hormones or hormone analogs, to each sow induction timing ovulation; And
Use deep intrauterine catheter or laparoscopic surgery one or more spermatid dosage that sow is inseminated, this one or more spermatid dosage being wherein administered to each sow altogether comprises and is less than 1x 10
9individual spermatid sum;
Reduce thus in this group or on this farm for the number of the required boar of breeding.
J1. the method as described in claim I or J, wherein superior in heredity boar comprises the boar relative to other boars in this group or on this farm with higher selection index.
K. for increasing a method for the rate of return (RMT) on swinery or farm, the method comprises:
The market condition stood based on this group or farm determines that male hog or female pigs cause the higher net profit of each pig;
Semen sample is collected from boar;
Described semen sample is divided and elects at least two spermatid subgroups as, if wherein this female pigs causes higher every pig net profit, then first subgroup of at least 80% is with X chromosome, if or this male hog causes higher every pig net profit, then first subgroup of at least 80% is with Y chromosome;
With the spermatid from described first subgroup, sow is inseminated; And
Offspring is produced from described sow.
K1. the method as described in K, wherein this male hog is hog.
K2. the method as described in K, wherein this female pigs is gilt.
K3. the method as described in K, be that male hog or female pigs cause this step of higher every pig net profit to comprise the character compared between male hog and female pigs under wherein determining the market condition stood this group or farm, wherein this character is selected from any one in the following: food conversion, body weight, average daily gain, carcass lean meat percentage, the loin degree of depth, the thickness of backfat, stomach fat thickness, carcass fat-free lean index, lean meat day increment, every pig feed cost and buccal fat iodine number.
As from can easily recognize above, basic conception of the present invention can embody in many ways.The present invention relates to the numerous and different embodiment using the spermatid of sex sorting to increase the genetic progress of a strain, include but not limited to best mode of the present invention.
Just because of this, to be disclosed by description or the specific embodiments of the invention of figure that the application encloses or table display or key element are not intended to be restrictive, but the numerous and different embodiment generally contained by the present invention or the example of equivalent contained about its any special key element.In addition, the specific descriptions of single embodiment of the present invention or key element may not describe all possible embodiment or key element clearly; Many replacement schemes are impliedly disclosed by description and accompanying drawing.
Claims (41)
1. increase a method for the genetic worth of pig, the method comprises the following steps:
Set up the multiple copulation hypotypes for a strain;
To determine for each in these copulation hypotypes be the percentage ratio of male filial generation or be the percentage ratio of female filial generation for each in these copulation hypotypes, and this percentage ratio will cause genetic worth increase in this strain relative to contrast;
Divided by spermatid sample from the male hog be in one of these copulation hypotypes and elect one or more spermatid subgroup as, the spermatid of in one of them spermatid subgroup at least 60% is with X chromosome or Y chromosome; And
The one or more female pigs inseminations be in one of these copulation hypotypes are made, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast with the spermatid of this subgroup.
2. the method for claim 1, is wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
3. the method for claim 1, wherein this male hog or this one or more female pigs are the members of hereditary core, daughter nucleus or multiplier.
4. the method for claim 1, wherein this male hog or this one or more female pigs are the members of hereditary core.
5. the method for claim 1, the percentage ratio of the spermatid wherein in the spermatid subgroup with X chromosome or Y chromosome is selected from lower group, and this group is made up of the following: at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and at least 100%.
6. the method for claim 1, wherein this strain comprises a paternal line or maternal.
7. the method for claim 1, wherein in this contrast, the filial generation of about 50% is male.
8. the method for claim 1, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
9. the method for claim 1, the classification of the male hog wherein in one or more copulation hypotype or the kind one or more features comprising genetic worth or age define.
10. the method for claim 1, the classification of the female pigs wherein in one or more copulation hypotype or the kind one or more features comprising genetic worth or parity define.
11. the method for claim 1, wherein this strain comprises a paternal line, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 10% to 35%.
12. methods as claimed in claim 11 are wherein that the percentage ratio of male filial generation is between about 15% to 30% for these product.
13. the method for claim 1, wherein this strain comprise one maternal, and be the percentage ratio of male filial generation for each in these copulation hypotypes relative to contrast by what determine to increase the genetic worth of this strain or be the percentage ratio of female filial generation for each in these copulation hypotypes, cause being the percentage ratio of male filial generation for these product between about 5% to 30%.
14. methods as claimed in claim 13 are wherein that the percentage ratio of male filial generation is between about 10% to 25% for these product.
15. the method for claim 1, wherein for each in these copulation hypotypes be male filial generation percentage ratio or for each in these copulation hypotypes be the percentage ratio of female filial generation between about 0 to 10% or between about 90% to 100%, and the filial generation of any generation is the member of daughter nucleus or multiplier.
16. the method for claim 1, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain or an only female pigs.
17. methods as claimed in claim 16, each of these copulation hypotypes wherein in multiple copulation hypotype comprises an only male hog from this strain and an only female pigs.
18. the method for claim 1, wherein these percentage ratios determined use randomness or Deterministic Methods to determine.
19. 1 kinds of methods increasing the genetic worth of pig, the method comprises the following steps:
Set up the multiple copulation hypotypes for a strain;
To determine for each in these copulation hypotypes be the percentage ratio of male filial generation or be the percentage ratio of female filial generation for each in these copulation hypotypes, and this percentage ratio will cause genetic worth increase in this strain relative to contrast;
Divided by spermatid sample from the male hog be in one of these copulation hypotypes and elect one or more spermatid subgroup as, the spermatid of in one of them spermatid subgroup at least 60% is with X chromosome or Y chromosome; And
The one or more ovum fertilizations from the one or more female pigs be in one of these copulation hypotypes are made, to realize being determined to increase the percentage ratio for male filial generation of genetic worth or the percentage ratio for female filial generation relative to contrast with the spermatid of this subgroup.
20. methods as claimed in claim 19, are wherein determined to increase the percentage ratio for male filial generation of genetic worth or for the percentage ratio of female filial generation is relative to contrasting the inbreeding be not increased in this strain.
21. methods as claimed in claim 19, wherein in this contrast, the filial generation of about 50% is male.
22. methods as claimed in claim 19, wherein in this contrast, needing all female pigs of copulation inseminates with unsorted semen sample.
23. 1 kinds of methods being increased in the genetic worth in paternal line, the method comprises the following steps:
Divided by one or more spermatid samples from the one or more male hogs in this paternal line and elect one or more spermatid subgroup as, in one of them subgroup, the spermatid of at least 60% is with X chromosome or Y chromosome; And
The one or more female pigs insemination in this paternal line is made, to realize being the percentage ratio of male filial generation for this paternal line between about 10% to 35% by this subgroup.
24. methods as claimed in claim 23 are wherein that the percentage ratio of male filial generation is between about 15% to 30% for this paternal line.
25. methods as claimed in claim 23, wherein this strain belongs to hereditary core, daughter nucleus or multiplier.
26. methods as claimed in claim 23, wherein this strain belongs to hereditary core.
27. methods as claimed in claim 23, the percentage ratio of the spermatid wherein in the spermatid subgroup with X chromosome or Y chromosome is selected from lower group, and this group is made up of the following: at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and at least 100%.
28. 1 kinds be increased in maternal in the method for genetic worth, the method comprises the following steps:
Divided by one or more spermatid samples of the one or more male hogs in maternal from this and elect one or more spermatid subgroup as, in one of them subgroup, the spermatid of at least 70% is with X chromosome or Y chromosome; And
One or more female pigs insemination in making this maternal by this subgroup, with realize between about 5% to 30% for this maternal percentage ratio for male filial generation.
29. methods as claimed in claim 28, wherein maternal for this is that the percentage ratio of male filial generation is between about 10% to 25%.
30. methods as claimed in claim 28, wherein this strain belongs to hereditary core, daughter nucleus or multiplier.
31. methods as claimed in claim 28, wherein this strain belongs to hereditary core.
32. methods as claimed in claim 28, the percentage ratio of the spermatid wherein in the spermatid subgroup with X chromosome or Y chromosome is selected from lower group, and this group is made up of the following: at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% and at least 100%.
33. the method for claim 1, the deep intrauterine catheter wherein making one or more female pigs inseminate to comprise and use and inserted by the film of one or more female pigs or pin use this subgroup to the reproductive tract of this one or more female pigs.
34. methods as claimed in claim 33, wherein use deep intrauterine catheter to use this subgroup to the reproductive tract of one or more female pigs and comprise and being inserted in one or more cornua uteri by described spermatid.
35. methods as claimed in claim 34, wherein said deep intrauterine catheter comprises a video camera or endoscope.
36. methods as claimed in claim 34, comprise the step made via radiography or fluoroscopy in the visual reproductive tract simultaneously inserting described sow of this deep intrauterine catheter further.
37. methods as claimed in claim 34, wherein this subgroup comprises 1x10
9individual or less spermatid.
38. methods as claimed in claim 37, wherein this subgroup is 300x10
6individual less spermatid, 150x10
6individual or less spermatid, 140x10
6individual or less spermatid, 100x10
6individual or less spermatid, 70x10
6individual or less spermatid, 50x10
6individual or less spermatid or 5x10
6individual or less spermatid.
39. methods as claimed in claim 33, are wherein used the pin that inserted by the film of this one or more female pigs to use this subgroup to the reproductive tract of this one or more female pigs and comprise and this subgroup being expelled in one or more fallopian tube of this one or more female pigs.
40. methods as claimed in claim 39, comprise further and make via a peritoneoscope or video camera the visual step of described pin that is inserted in described one or more fallopian tube.
41. methods as described in any of claims 39, wherein this subgroup comprises 1x10
6individual or less spermatid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310368889.8A CN116211532A (en) | 2013-03-15 | 2013-11-27 | Methods of increasing genetic value in male or female lines using sexed sperm |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US13/840,598 US9433195B2 (en) | 2012-06-06 | 2013-03-15 | Methods for increasing genetic progress in a line or breed of swine using sex-selected sperm cells |
| US13/840,598 | 2013-03-15 | ||
| PCT/US2013/072272 WO2014143223A1 (en) | 2013-03-15 | 2013-11-27 | Methods for use of sex sorted semen to improve genetic management in swine |
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| CN202310368889.8A Division CN116211532A (en) | 2013-03-15 | 2013-11-27 | Methods of increasing genetic value in male or female lines using sexed sperm |
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| CN202310368889.8A Pending CN116211532A (en) | 2013-03-15 | 2013-11-27 | Methods of increasing genetic value in male or female lines using sexed sperm |
| CN201380074208.1A Pending CN105025905A (en) | 2013-03-15 | 2013-11-27 | Methods for use of sex sorted semen to improve genetic management in swine |
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| CN202310368889.8A Pending CN116211532A (en) | 2013-03-15 | 2013-11-27 | Methods of increasing genetic value in male or female lines using sexed sperm |
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| Country | Link |
|---|---|
| EP (1) | EP2968421A4 (en) |
| CN (2) | CN116211532A (en) |
| BR (1) | BR112015023623A2 (en) |
| CA (1) | CA2904193C (en) |
| CL (1) | CL2015002365A1 (en) |
| HK (1) | HK1217284A1 (en) |
| MX (2) | MX2020011408A (en) |
| WO (1) | WO2014143223A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109195506A (en) * | 2016-05-25 | 2019-01-11 | 穆格公司 | Sensing system for more lumen tubes |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9433195B2 (en) * | 2012-06-06 | 2016-09-06 | Inguran, Llc | Methods for increasing genetic progress in a line or breed of swine using sex-selected sperm cells |
Citations (3)
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| WO2001051612A1 (en) * | 2000-01-14 | 2001-07-19 | Istituto Sperimentale Italiano 'lazzaro Spallanzani' | Process for the production of non-human embryos of high-genetic value and of predetermined sex |
| CN1433267A (en) * | 2000-05-09 | 2003-07-30 | Xy公司 | High purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa |
| CN1633259A (en) * | 2000-06-12 | 2005-06-29 | Xy公司 | Integrated herd management system utilizing isolated populations of X-chromosome bearing and Y-chromosome bearing spermatozoa |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9888990B2 (en) * | 2012-06-06 | 2018-02-13 | Inguran, Llc | Methods for use of sex sorted semen to improve genetic management in swine |
| US9433195B2 (en) * | 2012-06-06 | 2016-09-06 | Inguran, Llc | Methods for increasing genetic progress in a line or breed of swine using sex-selected sperm cells |
-
2013
- 2013-11-27 BR BR112015023623A patent/BR112015023623A2/en not_active Application Discontinuation
- 2013-11-27 CA CA2904193A patent/CA2904193C/en active Active
- 2013-11-27 CN CN202310368889.8A patent/CN116211532A/en active Pending
- 2013-11-27 CN CN201380074208.1A patent/CN105025905A/en active Pending
- 2013-11-27 MX MX2020011408A patent/MX2020011408A/en unknown
- 2013-11-27 MX MX2015012648A patent/MX358745B/en active IP Right Grant
- 2013-11-27 EP EP13877728.9A patent/EP2968421A4/en active Pending
- 2013-11-27 WO PCT/US2013/072272 patent/WO2014143223A1/en active Application Filing
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001051612A1 (en) * | 2000-01-14 | 2001-07-19 | Istituto Sperimentale Italiano 'lazzaro Spallanzani' | Process for the production of non-human embryos of high-genetic value and of predetermined sex |
| CN1433267A (en) * | 2000-05-09 | 2003-07-30 | Xy公司 | High purity X-chromosome bearing and Y-chromosome bearing populations of spermatozoa |
| CN1633259A (en) * | 2000-06-12 | 2005-06-29 | Xy公司 | Integrated herd management system utilizing isolated populations of X-chromosome bearing and Y-chromosome bearing spermatozoa |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109195506A (en) * | 2016-05-25 | 2019-01-11 | 穆格公司 | Sensing system for more lumen tubes |
| CN109195506B (en) * | 2016-05-25 | 2021-08-24 | 穆格公司 | Sensing system for multi-lumen tubing |
| US11097050B2 (en) | 2016-05-25 | 2021-08-24 | Moog Inc. | Sensing system for multiple lumen tubing |
Also Published As
| Publication number | Publication date |
|---|---|
| MX358745B (en) | 2018-08-31 |
| HK1217284A1 (en) | 2017-01-06 |
| EP2968421A1 (en) | 2016-01-20 |
| MX2020011408A (en) | 2021-09-15 |
| CA2904193A1 (en) | 2014-09-18 |
| CL2015002365A1 (en) | 2016-03-04 |
| CA2904193C (en) | 2023-04-04 |
| MX2015012648A (en) | 2016-02-18 |
| BR112015023623A2 (en) | 2018-05-08 |
| CN116211532A (en) | 2023-06-06 |
| WO2014143223A1 (en) | 2014-09-18 |
| EP2968421A4 (en) | 2016-09-21 |
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